TRANSPOSON SYSTEM AND METHODS OF USE

RELATED APPLICATIONS

[01] This application claims the benefit of provisional application USSN 62/552,861, filed August 31, 2017, USSN 62/558,286, filed September 13, 2017 and USSN 62/608,546, filed December 20, 2017, the contents of each of which are herein incorporated by reference in their entirety.

INCORPORATION OF SEQUENCE LISTING

[02] The contents of the text file named "POTH-029/001WO_SeqList.txt," which was created on August 31, 2018 and is 44,366 KB in size, are hereby incorporated by reference in their entirety.

FIELD OF THE DISCLOSURE

[03] The present invention is directed to compositions and methods for targeted gene modification.

BACKGROUND

[04] Ex vivo genetic modification of non-transformed primary human T lymphocytes using non-viral vector-based gene transfer delivery systems has been extremely difficult. As a result, most groups have generally used viral vector-based transduction such as retrovirus, including lentivirus. A number of non-viral methods have been tested and include antibody- targeted liposomes, nanoparticles, aptamer siRNA chimeras, electroporation, nucleofection, lipofection, and peptide transduction. Overall, these approaches have resulted in poor transfection efficiency, direct cell toxicity, or a lack of experimental throughput.

[05] The use of plasmid vectors for genetic modification of human lymphocytes has been limited by low efficiency using currently available plasmid transfection systems and by the toxicity that many plasmid transfection reagents have on these cells. There is a long-felt and unmet need for a method of nonviral gene modification in immune cells.

SUMMARY

[06] When compared with viral transduction of immune cells, such as T lymphocytes, delivery of transgenes via DNA transposons, such as piggyBac and Sleeping Beauty, offers significant advantages in ease of use, ability to delivery much larger cargo, speed to clinic and cost of production. The piggyBac DNA transposon, in particular, offers additional advantages in giving long-term, high-level and stable expression of transgenes, and in being significantly less mutagenic than a retrovirus, being non-oncogenic and being fully reversible. Previous attempts to use DNA transposons to deliver transgenes to T cells have been unsuccessful at generating commercially viable products or manufacturing methods because the previous methods have been inefficient. For example, the poor efficiency demonstrated by previous methods of using DNA transposons to deliver transgenes to T cells has resulted in the need for prolonged expansion ex vivo. Previous unsuccessful attempts by others to solve this problem have all focused on increasing the amount of DNA transposon delivered to the immune cell, which has been a strategy that worked well for non-immune cells. This disclosure demonstrates that increasing the amount of DNA transposon makes the efficiency problem worse in immune cells by increasing DNA-mediated toxicity. To solve this problem, counterintuitively, the methods of the disclosure decrease the amount of DNA delivered to the immune cell. Using the methods of the disclosure, the data provided herein demonstrate not only that decreasing the amount of DNA transposon introduced into the cell increased viability but also that this method increased the percentage of cells that harbored a transposition event, resulting in a viable commercial process and a viable commercial product. Thus, the methods of the disclosure demonstrate success where others have failed.

[07] The disclosure provides a nonviral method for the ex-vivo genetic modification of an immune cell or an immune cell precursor comprising delivering to the immune cell or the immune cell precursor, (a) a nucleic acid or amino acid sequence comprising a sequence encoding a transposase enzyme and (b) a recombinant and non-naturally occurring DNA sequence comprising a DNA sequence encoding a transposon. In certain embodiments, the method further comprises the step of stimulating the immune cell or the immune cell precursor with one or more cytokine(s).

[08] In certain embodiments of the methods of the disclosure, the sequence encoding a transposase enzyme is an mRNA sequence. The mRNA sequence encoding a transposase enzyme may be produced in vitro.

[09] In certain embodiments of the methods of the disclosure, the sequence encoding a transposase enzyme is a DNA sequence. The DNA sequence encoding a transposase enzyme may be produced in vitro. The DNA sequence may be a cDNA sequence.

[010] In certain embodiments of the methods of the disclosure, the sequence encoding a transposase enzyme is an amino acid sequence. The amino acid sequence encoding a transposase enzyme may be produced in vitro. A protein Super piggybac transposase (SPB) may be delivered following pre-incubation with transposon DNA.

[Oil] In certain embodiments of the methods of the disclosure, the delivering step comprises electroporation or nucleofection of the immune cell or the immune cell precursor.

[012] In certain embodiments of the methods of the disclosure, the method further comprises the step of stimulating the immune cell or the immune cell precursor with one or more cytokines. In certain embodiments, the step of stimulating the immune cell or the immune cell precursor with one or more cytokine(s) occurs following the delivering step. Alternatively, or in addition, in certain embodiments, the step of stimulating the immune cell or the immune cell precursor with one or more cytokine(s) occurs prior to the delivering step. In certain embodiments, the one or more cytokine(s) comprise(s) IL-2, IL-21, IL-7 and/or IL- 15.

[013] In certain embodiments of the methods of the disclosure, the immune cell or the immune cell precursor is an autologous immune cell or immune cell precursor. The immune cell or immune cell precursor may be a human immune cell, a human immune cell precursor, an autologous immune cell, and/or an autologous immune cell precursor. The immune cell may be derived from a non-autologous source, including, but not limited to a primary cell, a cultured cell or cell line, an embryonic or adult stem cell, an induced pluripotent stem cell or a transdifferentiated cell. The immune cell may have been previously genetically modified or derived from a cell or cell line that has been genetically modified. The immune cell may be modified or may be derived from a cell or cell line that has been modified to suppress one or more apoptotic pathways. The immune cell may be modified or may be derived from a cell or cell line that has been modified to be "universally" allogenic by a majority of recipients in the context, for example, of a therapy involving an adoptive cell transfer.

[014] In certain embodiments of the methods of the disclosure, the immune cell is an activated immune cell.

[015] In certain embodiments of the methods of the disclosure, the immune cell is a resting immune cell.

[016] In certain embodiments of the methods of the disclosure, the immune cell is a T- lymphocyte. In certain embodiments, the T-lymphocyte is an activated T-lymphocyte. In certain embodiments, the T-lymphocyte is a resting T-lymphocyte.

[017] In certain embodiments of the methods of the disclosure, the immune cell is a Natural Killer (NK) cell. [018] In certain embodiments of the methods of the disclosure, the immune cell is a Cytokine-induced Killer (CIK) cell.

[019] In certain embodiments of the methods of the disclosure, the immune cell is a Natural Killer T (NKT) cell.

[020] In certain embodiments of the methods of the disclosure, the immune cell is isolated or derived from a human.

[021] In certain embodiments of the methods of the disclosure, the immune cell precursor is a stem cell or stem-like cell capable of differentiation into an immune cell. In some embodiments, the immune cell precursor is a hematopoietic stem cell (HSC). In some embodiments, the immune cell precursor is a primitive hematopoietic stem cell. In some embodiments, the immune cell precursor is a human HSC or human primitive HSC.

[022] In certain embodiments of the methods of the disclosure, the method further comprising the step of differentiating the immune cell precursor into an immune cell. In some embodiments, the immune cell is a T lymphocyte (T cell), a B lymphocyte (B cell), a Natural Killer (NK) cell, or a Cytokine-induced Killer (CIK) cell.

[023] In certain embodiments of the methods of the disclosure, the immune cell is isolated or derived from a non-human mammal. In certain embodiments, the non-human mammal is a rodent, a rabbit, a cat, a dog, a pig, a horse, a cow, or a camel. In certain embodiments, the immune cell is isolated or derived from a non-human primate.

[024] In certain embodiments of the methods of the disclosure, the mRNA sequence encoding the transposase enzyme is produced in vitro.

[025] In certain embodiments, the transposon is a piggyBac transposon or a piggyBac-like transposon. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac transposase. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac- like transposon, the transposase is a piggyBac-like transposase.

[026] In certain embodiments, the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 14487. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™ or a Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a Super piggyBac™ (SPB) transposase, the sequence encoding the transposase is an mRNA sequence.

[027] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme. The piggyBac (PB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV

541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID

NO: 14487) .

[028] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at one or more of positions 30, 165, 282, or 538 of the sequence:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV

541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID

NO: 14487) .

[029] In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at two or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at three or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at each of the following positions 30, 165, 282, and 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 165 of the sequence of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G). In certain embodiments, the amino acid substitution at position 282 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 538 of the sequence of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N).

[030] In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) transposase enzyme. In certain embodiments, the Super piggyBac™ (SPB) transposase enzymes of the disclosure may comprise or consist of the amino acid sequence of the sequence of SEQ ID NO: 14487 wherein the amino acid substitution at position 30 is a substitution of a valine (V) for an isoleucine (I), the amino acid substitution at position 165 is a substitution of a serine (S) for a glycine (G), the amino acid substitution at position 282 is a substitution of a valine (V) for a methionine (M), and the amino acid substitution at position 538 is a substitution of a lysine (K) for an asparagine (N). In certain embodiments, the Super piggyBac™ (SPB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV

541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID

14484) .

[031] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 3, 46, 82, 103, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 258, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 486, 503, 552, 570 and 591 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 46, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 485, 503, 552 and 570. In certain embodiments, the amino acid substitution at position 3 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for a serine (S). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an alanine (A). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 82 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 119 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for an arginine (R). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) a cysteine (C). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a

phenylalanine (F). In certain embodiments, the amino acid substitution at position 185 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 187 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for an alanine (A). In certain embodiments, the amino acid substitution at position 200 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a phenylalanine (F).In certain embodiments, the amino acid substitution at position 207 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a valine (V). In certain embodiments, the amino acid substitution at position 209 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a valine (V). In certain embodiments, the amino acid substitution at position 226 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a methionine (M). In certain embodiments, the amino acid substitution at position 235 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a leucine (L). In certain embodiments, the amino acid substitution at position 240 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 241 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 243 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a proline (P). In certain embodiments, the amino acid substitution at position 258 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a proline (P). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine for a proline (P). In certain embodiments, the amino acid substitution at position 315 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for an arginine (R).In certain embodiments, the amino acid substitution at position 319 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a threonine (T). In certain embodiments, the amino acid substitution at position 327 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 328 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a cysteine (C). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 421 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for the aspartic acid (D). In certain embodiments, the amino acid substitution at position 436 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a valine (V). In certain embodiments, the amino acid substitution at position 456 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a methionine (M). In certain embodiments, the amino acid substitution at position 470 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 485 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a serine (S). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a methionine (M). In certain embodiments, the amino acid substitution at position 552 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a glutamine (Q). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a glutamine (Q).

[032] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at two, three, four, five, six or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 194 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 372 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for an arginine (R). In certain embodiments, the amino acid substitution at position 375 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a lysine (K). In certain embodiments, the amino acid substitution at position 450 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for an aspartic acid (D). In certain embodiments, the amino acid substitution at position 509 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a serine (S). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487. In certain embodiments, including those embodiments wherein the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, the piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 372, 375 and 450 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, and a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487. In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487 and a substitution of an asparagine (N) for an aspartic acid (D) at position 450 of SEQ ID NO: 14487.

[033] In certain embodiments of the methods of the disclosure, the transposase enzyme is a

Super piggyBac™ (SPB) transposase enzyme. The Super piggyBac (PB) transposase enzyme may comprise or consist of an amino acid sequence at least 75% identical to:

MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEEAFIDEVHEVQPTS

SGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKSTRRSRVSALNIV RS

QRGPTPJVICRNIYDPLLCFia.FFTDEIISEIVKWTOAEISLKRRESMTSATFRDT NEDEI

YAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKSIR

PTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRVYIPNKPS KY

GIIQLMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSCRNITCDN

WFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDGPLTLV SY

KPKPAIGVIVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRI

TORWPMALLYGMINIACINSFIIYSHNVSSKGEKVQSPJ KFMRNLYMSLTSSFMRKR

LEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTTCTTCPSiaPJIKANASC KK

CKKVICREHNIDMCQSCF (SEQ ID NO: 14484).

[034] In certain embodiments of the methods of the disclosure, the transposon is a Sleeping Beauty transposon. In certain embodiments of the methods of the disclosure, the transposase enzyme is a Sleeping Beauty transposase enzyme (see, for example, US Patent No. 9,228,180, the contents of which are incorporated herein in their entirety). In certain embodiments, the Sleeping Beauty transposase is a hyperactive Sleeping Beauty (SB100X) transposase. In certain embodiments, the Sleeping Beauty transposase enzyme comprises an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGKSKEISQD LRKKIVDLHK SGSSLGAISK RLKVPRSSVQ TIVRKYKHHG TTQPSYRSGR

61 RRVLSPRDER TLVRKVQINP RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGRSARKK

121 PLLQNRHKKA RLRFATAHGD KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN

181 TIPTVKHGGG SIMLWGCFAA GGTGALHKID GIMRKENYVD ILKQHLKTSV RKLKLGRK V

241 FQMDNDPKHT SKWAKWLKD NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL

301 HQLCQEEWAK IHPTYCGKLV EGYPKRLTQV KQFKGNATKY (SEQ ID NO: 14485). In certain embodiments, including those wherein the Sleeping Beauty transposase is a hyperactive Sleeping Beauty (SB100X) transposase, the Sleeping Beauty transposase enzyme comprises an amino acid sequence at least at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGKSKEISQD LRKRIVDLHK SGSSLGAISK RLAVPRSSVQ TIVRKYKHHG TTQPSYRSGR

61 RRVLSPRDER TLVRKVQINP RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGHSARKK

121 PLLQNRHKKA RLRFATAHGD KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN

181 TIPTVKHGGG SIMLWGCFAA GGTGALHKID GIMDAVQYVD ILKQHLKTSV RKLKLGRKWV

241 FQHDNDPKHT SKWAKWLKD NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL

301 HQLCQEEWAK IHPNYCGKLV EGYPKRLTQV KQFKGNATKY (SEQ ID NO: 14486).

[035] In certain embodiments of the methods of the disclosure, the transposon is a Helraiser transposon. In certain embodiments of the Helraiser transposon sequence, the transposase is flanked by left and right terminal sequences termed LTS and RTS. In certain embodiments, these sequences terminate with a conserved 5'-TC/CTAG-3' motif. In certain embodiments, a 19 bp palindromic sequence with the potential to form the hairpin termination structure is located 11 nucleotides upstream of the RTS and comprises the sequence

GTGCACGAATTTCGTGCACCGGGCCACTAG (SEQ ID NO: 14500).

[036] In certain embodiments of the methods of the disclosure, and, in particular those embodiments wherein the transposon is a Helraiser transposon, the transposase enzyme is a Helitron transposase enzyme. In certain embodiments, the Helitron transposase enzyme of the disclosure comprises an amino acid sequence comprising:

1 MSKEQLLIQR SSAAERCRRY RQKMSAEQRA SDLERRRRLQ QNVSEEQLLE KRRSEAEKQR

61 RHRQKMSKDQ RAFEVERRRW RRQNMSREQS STSTTNTGRN CLLSKNGVHE DAILEHSCGG

121 MTVRCEFCLS LNFSDEKPSD GKFTRCCSKG KVCPNDIHFP DYPAYLKRLM TNEDSDSKNF

181 MENIRSINSS FAFASMGANI ASPSGYGPYC FRIHGQVYHR TGTLHPSDGV SRKFAQLYIL

241 DTAEATSKRL AMPENQGCSE RLMININNLM HEINELTKSY KMLHEVEKEA QSEAAAKGIA

301 PTEVTMAIKY DRNSDPGRYN SPRVTEVAVI FRNEDGEPPF ERDLLIHCKP DPNNPNATKM

361 KQISILFPTL DAMTYPILFP HGEKGWGTDI ALRLRDNSVI DNNTRQNVRT RVTQMQYYGF

421 HLSVRDTFNP ILNAGKLTQQ FIVDSYSKME ANRINFIKAN QSKLRVEKYS GLMDYLKSRS

481 ENDNVPIGKM IILPSSFEGS PRNMQQRYQD AMAIVTKYGK PDLFITMTCN PKWADITNNL

541 QRWQKVENRP DLVARVFNIK LNALLNDICK FHLFGKVIAK IHVIEFQKRG LPHAHILLIL

601 DSESKLRSED DIDRIVKAEI PDEDQCPRLF QIVKSNMVHG PCGIQNPNSP CMENGKCSKG

661 YPKEFQNATI GNIDGYPKYK RRSGSTMSIG NKWDNTWIV PYNPYLCLKY NCHINVEVCA

721 SIKSVKYLFK YIYKGHDCAN IQISEKNIIN HDEVQDFIDS RYVSAPEAVW RLFAMRMHDQ

781 SHAITRLAIH LPNDQNLYFH TDDFAEVLDR AKRHNSTLMA WFLLNREDSD ARNYYYWEIP

841 QHYVFNNSLW TKRRKGGNKV LGRLFTVSFR EPERYYLRLL LLHVKGAISF EDLRTVGGVT

901 YDTFHEAAKH RGLLLDDTIW KDTIDDAIIL NMPKQLRQLF AYICVFGCPS AADKLWDENK

961 SHFIEDFCWK LHRREGACV CEMHALNEIQ EVFTLHGMKC SHFKLPDYPL LMNANTCDQL

1021 YEQQQAEVLI NSLNDEQLAA FQTITSAIED QTVHPKCFFL DGPGGSGKTY LYKVLTHYIR

1081 GRGGTVLPTA STGIAANLLL GGRTFHSQYK LPIPLNETSI SRLDIKSEVA KTIKKAQLLI 1141 IDECTMASSH AINAIDRLLR EIMNLNVAFG GKVLLLGGDF RQCLSIVPHA MRSAIVQTSL

1201 KYCNVWGCFR KLSLKTNMRS EDSAYSEWLV KLGDGKLDSS FHLGMDI IEI PHEMICNGSI

1261 IEATFGNSIS IDNIKNISKR AILCPKNEHV QKLNEEILDI LDGDFHTYLS DDSIDSTDDA

1321 EKENFPIEFL NSITPSGMPC HKLKLKVGAI IMLLRNLNSK WGLCNGTRFI IKRLRPNI IE

1381 AEVLTGSAEG EWLIPRIDL SPSDTGLPFK LIRRQFPVMP AFAMTINKSQ GQTLDRVGIF

1441 LPEPVFAHGQ LYVAFSRVRR ACDVKVKW TSSQGKLVKH SESVFTLNW YREILE (SEQ ID

NO: 14501).

[037] In certain embodiments of the methods of the disclosure, the transposon is a Tol2 transposon.

[038] In certain embodiments of the methods of the disclosure, and, in particular those embodiments wherein the transposon is a Tol2 transposon, the transposase enzyme is a Tol2 transposase enzyme. In certain embodiments, the Tol2 transposase enzyme of the disclosure comprises an amino acid sequence comprising:

1 MEEVCDSSAA ASSTVQNQPQ DQEHPWPYLR EFFSLSGVNK DSFKMKCVLC LPLNKEISAF

61 KSSPSNLRKH IERMHPNYLK NYSKLTAQKR KIGTSTHASS SKQLKVDSVF PVKHVSPVTV

121 NKAILRYIIQ GLHPFSTVDL PSFKELISTL QPGISVITRP TLRSKIAEAA LIMKQKVTAA

181 MSEVEWIATT TDCWTARRKS FIGVTAHWIN PGSLERHSAA LACKRLMGSH TFEVLASAMN

241 DIHSEYEIRD KWCTTTDSG SNFMKAFRVF GVENNDIETE ARRCESDDTD SEGCGEGSDG

301 VEFQDASRVL DQDDGFEFQL PKHQKCACHL LNLVSSVDAQ KALSNEHYKK LYRSVFGKCQ

361 ALWNKSSRSA LAAEAVESES RLQLLRPNQT RWNSTFMAVD RILQICKEAG EGALRNICTS

421 LEVPMFNPAE MLFLTEWANT MRPVAKVLDI LQAETNTQLG WLLPSVHQLS LKLQRLHHSL

481 RYCDPLVDAL QQGIQTRFKH MFEDPEI IAA AILLPKFRTS WTNDETIIKR GMDYIRVHLE

541 PLDHKKELAN SSSDDEDFFA SLKPTTHEAS KELDGYLACV SDTRESLLTF PAICSLSIKT

601 NTPLPASAAC ERLFSTAGLL FSPKRARLDT NNFENQLLLK LNLRFYNFE (SEQ ID NO:

14502).

[039] In certain embodiments of the methods of the disclosure, the piggyBac-like transposon comprises an amino acid sequence having at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or any percentage in between of identity to the amino acid sequence of SEQ ID NO: 14487.

[040] In certain embodiments of the methods of the disclosure, a vector comprises the recombinant and non-naturally occurring DNA sequence encoding the transposon. In some embodiments, the vector comprises any form of DNA and wherein the vector comprises at least 100 nucleotides (nts), 500 nts, 1000 nts, 1500 nts, 2000 nts, 2500 nts, 3000 nts, 3500 nts, 4000 nts, 4500 nts, 5000 nts, 6500 nts, 7000 nts, 7500 nts, 8000 nts, 8500 nts, 9000 nts, 9500 nts, 10,000 nts or any number of nucleotides in between. In some embodiments, the vector comprises single-stranded or double-stranded DNA. In some embodiments, the vector comprises circular DNA. In some embodiments, the vector is a plasmid vector. In some embodiments, the vector is a nanoplasmid vector. In some embodiments, the vector is a minicircle. In some embodiments, the vector comprises linear or linearized DNA. In some embodiments, the linear or linearized DNA is produced in vitro. In some embodiments, the linear or linearized DNA is a product of a restriction digest of a circular DNA. In some embodiments, the circular DNA is a plasmid vector, a nanoplasmid vector or a minicircle DNA vector. In some embodiments, the linear or linearized DNA is a product of a polymerase chain reaction (PCR). In some embodiments, the vector is a double-stranded doggybone™ DNA sequence. In some embodiments, the doggybone™ DNA sequence is produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, poly -A tail and telomeric ends.

[041] In certain embodiments of the methods of the disclosure, the immune cell or the immune cell precursor is isolated or derived from a human. In certain embodiments, the immune cell or the immune cell precursor is isolated or derived from a non-human mammal. In certain embodiments, the non-human mammal is a rodent, a rabbit, a cat, a dog, a pig, a horse, a cow, a camel or a primate.

[042] In certain embodiments of the methods of the disclosure, the recombinant and non- naturally occurring DNA sequence encoding a transposon further comprises a sequence encoding a chimeric antigen receptor or a portion thereof. In certain embodiments, the chimeric antigen receptor (CAR) comprises (a) an ectodomain comprising an antigen recognition region, (b) a transmembrane domain, and (c) an endodomain comprising at least one costimulatory domain. In certain embodiments, the antigen recognition region comprises one or more of an antibody or a fragment thereof; a single chain antibody (scFv), a single domain antibody, an antibody mimetic, a protein scaffold, a Centyrin, a VHH, and a VH.

[043] Chimeric antigen receptors (CARs) of the disclosure may comprise (a) an ectodomain comprising an antigen recognition region, (b) a transmembrane domain, and (c) an endodomain comprising at least one costimulatory domain. In certain embodiments, the ectodomain may further comprise a signal peptide. Alternatively, or in addition, in certain embodiments, the ectodomain may further comprise a hinge between the antigen recognition region and the transmembrane domain. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD2, CD35, CD3s, CD3y, ΟΌ3ζ, CD4, CD8a, CD19, CD28, 4-1BB or GM-CSFR signal peptide. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD 8a signal peptide. In certain embodiments, the transmembrane domain may comprise a sequence encoding a human CD2, CD35, CD3s, CD3y, CO3C,, CD4, CD8a, CD19, CD28, 4-1BB or GM-CSFR transmembrane domain. In certain embodiments of the CARs of the disclosure, the transmembrane domain may comprise a sequence encoding a human CD8a transmembrane domain. In certain

embodiments of the CARs of the disclosure, the endodomain may comprise a human CD3ζ endodomain. In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a human 4-1BB, CD28, CD40, ICOS, MyD88, OX-40 intracellular segment, or any combination thereof. In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a CD28 and/or a 4-1BB costimulatory domain. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8a, IgG4, and/or CD4 sequence. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD 8 a sequence.

[044] In certain embodiments of the methods of the disclosure, the recombinant and non- naturally occurring DNA sequence encoding a transposon further comprises a sequence encoding a chimeric antigen receptor or a portion thereof. The portion of the sequence encoding a chimeric antigen receptor may encode an antigen recognition region. The antigen recognition region may comprise one or more complementarity determining region(s). The antigen recognition region may comprise an antibody, an antibody mimetic, a protein scaffold or a fragment thereof. In certain embodiments, the antibody is a chimeric antibody, a recombinant antibody, a humanized antibody or a human antibody. In certain embodiments, the antibody is affinity -tuned. Nonlimiting examples of antibodies of the disclosure include a single-chain variable fragment (scFv), a VHH, a single domain antibody (sdAB), a small modular immunopharmaceutical (SMIP) molecule, or a nanobody. In certain embodiments, the VHH is camelid. Alternatively, or in addition, in certain embodiments, the VHH is humanized. Nonlimiting examples of antibody fragments of the disclosure include a complementary determining region, a variable region, a heavy chain, a light chain, or any combination thereof. Nonlimiting examples of antibody mimetics of the disclosure include an affibody, an afflilin, an affimer, an affitin, an alphabody, an anticalin, and avimer, a DARPin, a Fynomer, a Kunitz domain peptide, or a monobody. Nonlimiting examples of protein scaffolds of the disclosure include a Centyrin.

[045] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 10.0 μg per 100 of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 100 μg/mL.

[046] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 7.5 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 75 μg/mL.

[047] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 6.0 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 60 μg/mL. In certain embodiments, the transposase is a Sleeping Beauty transposase. In certain embodiments, the Sleeping Beauty transposase is a Sleeping Beauty 100X (SB100X) transposase.

[048] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 5.0 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 50 μg/mL.

[049] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 2.5 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 25 μg/mL.

[050] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 1.67 μg per 100 of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 16.7 μg/mL. In certain embodiments, the transposase is a Super piggyBac (PB) transposase. In certain embodiments, the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 14487.

[051] In certain embodiments of the methods of the disclosure, the transposase is a piggyBac transposase. In certain embodiments, the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 14487. In certain embodiments, the piggyBac transposase is a hyperactive variant and wherein the hyperactive variant comprises an amino acid substitution at one or more of positions 30, 165, 282 and 538 of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I) (I30V). In certain embodiments, the amino acid substitution at position 165 of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G) (G165S). In certain embodiments, the amino acid substitution at position 282 of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M) (M282V). In certain embodiments, the amino acid substitution at position 538 of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N) (N538K).

[052] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and (b) wherein an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 1.67 μg per 100 μΐ. of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 16.7 μg/mL. In certain embodiments, the transposase is a Super piggyBac (PB) transposase. In certain embodiments, the Super piggyBac (PB) transposase enzyme comprises an amino acid sequence at least 75% identical to: MGSSLDDEHILSALLQSDDELVGEDSDSEVSDHVSEDDVQSDTEEAFIDEVHEVQPTS

SGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKSTRRSRVSALNIV RS

QRGPTPJVlCRNIYDPLLCFKLFFTDEnSEIVKWTNAEISLKRRESMTSATFRDTNE DEI

YAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKSIR

PTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRVYIPNKPS KY

GIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSCRNITCDN

WFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLK SRSRPVGTSMFCFDGPLTLVSY

KPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRK

TORWPMALLYGMINIACINSFnYSHNVSSKGEKVQSPJ KFMRNLYMSLTSSFMRKR

LEAPTLKRYLRDNISNILPKEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCK K

CKKVICREHNIDMCQSCF (SEQ ID NO: 14484).

[053] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 0.55 μg per 100 of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 5.5 μg/mL.

[054] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 0.19 μg per 100 μΐ. of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.9 μg/mL.

[055] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a DNA sequence, and an amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon is equal to or less than 0.10 μg per 100 μΐ. of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposase enzyme and an amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.0 μg/mL.

[056] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 10.0 μg per 100 of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 100 μg/mL.

[057] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 7.5 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 75 μg/mL.

[058] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 6.0 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 60 μg/mL. In certain embodiments, the transposase is a Sleeping Beauty transposase. In certain embodiments, the Sleeping Beauty transposase is a Sleeping Beauty 100X (SB100X) transposase.

[059] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 5.0 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 50 μg/mL.

[060] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 2.5 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 25 μg/mL.

[061] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 1.67 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 16.7 μg/mL. In certain embodiments, the transposase is a Super piggyBac (PB) transposase.

[062] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 0.55 μg per 100 of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 5.5 μg/mL.

[063] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 0.19 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.9 μg/mL.

[064] In certain embodiments of the methods of the disclosure, the nucleic acid sequence encoding the transposase enzyme is a RNA sequence, and an amount of the DNA sequence encoding the transposon is equal to or less than 0.1 μg per 100 μί of an electroporation or nucleofection reaction. In certain embodiments, a concentration of the amount of the DNA sequence encoding the transposon in the electroporation or nucleofection reaction is equal to or less than 1.0 μg/mL.

[065] The disclosure provides an immune cell modified according to the method of the disclosure. The immune cell may be a T-lymphocyte, a Natural Killer (NK) cell, a Cytokine- induced Killer (CIK) cell or a Natural Killer T (NKT) cell. The immune cell may be further modified by a second gene editing tool, including, but not limited to those gene editing tools comprising an endonuclease operably-linked to either a Cas9 or a TALE sequence. In certain embodiments of the second gene editing tool, the endonuclease is operably-linked to either a Cas9 or a TALE sequence covalently. In certain embodiments of the second gene editing tool, the endonuclease is operably-linked to either a Cas9 or a TALE sequence non- covalently. In certain embodiments, the endonuclease comprises a Clo051 domain. In certain embodiments, Clo051 domain comprises a sequence of

EGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLFEMKVLELLVNEYGFKGRH LGGSRKPDGIVYSTTLEDNFGIIVDTKAYSEGYSLPISQADEMERYVRENSNRDEEVN PNKWWENFSEEVKKYYFVFISGSFKGKFEEQLRRLSMTTGVNGSAVNVVNLLLGAE KIRSGEMTIEELERAMFN SEFILKY (SEQ ID NO: 14503).

[066] In certain embodiments, the Cas9 is an inactivated Cas9 (dCas9). In certain embodiments, the inactivated Cas9 is isolated or derived from Staphylococcus aureus and comprises DIOA and N580A within the catalytic site. In certain embodiments, the Cas9 is a small and inactivated Cas9 (dSaCas9). In certain embodiments, the dSaCas9 comprises the amino acid sequence of

1 MKRNYILGLA IGITSVGYGI IDYETRDVID AGVRLFKEAN VENNEGRRSK RGARRLKRRR

61 RHRIQRVKKL LFDYNLLTDH SELSGINPYE ARVKGLSQKL SEEEFSAALL HLAKRRGVHN

121 VNEVEEDTGN ELSTKEQISR NSKALEEKYV AELQLERLKK DGEVRGSINR FKTSDYVKEA

181 KQLLKVQKAY HQLDQSFIDT YIDLLETRRT YYEGPGEGSP FGWKDIKEWY EMLMGHCTYF

241 PEELRSVKYA YNADLYNALN DLNNLVITRD ENEKLEYYEK FQIIENVFKQ KKKPTLKQIA

301 KEILVNEEDI KGYRVTSTGK PEFTNLKVYH DIKDITARKE IIENAELLDQ IAKILTIYQS

361 SEDIQEELTN LNSELTQEEI EQISNLKGYT GTHNLSLKAI NLILDELWHT NDNQIAIFNR

421 LKLVPKKVDL SQQKEIPTTL VDDFILSPW KRSFIQSIKV INAI IKKYGL PNDIIIELAR

481 EKNSKDAQKM INEMQKRNRQ TNERIEEIIR TTGKENAKYL IEKIKLHDMQ EGKCLYSLEA

541 IPLEDLLNNP FNYEVDHI IP RSVSFDNSFN NKVLVKQEEA SKKGNRTPFQ YLSSSDSKIS

601 YETFKKHILN LAKGKGRISK TKKEYLLEER DINRFSVQKD FINRNLVDTR YATRGLMNLL

661 RSYFRV NLD VKVKSINGGF TSFLRRKWKF KKERNKGYKH HAEDALI IAN ADFIFKEWKK

721 LDKAKKVMEN QMFEEKQAES MPEIETEQEY KEIFITPHQI KHIKDFKDYK YSHRVDKKPN

781 RELINDTLYS TRKDDKGNTL IV NLNGLYD KDNDKLKKLI NKSPEKLLMY HHDPQTYQKL

841 KLIMEQYGDE KNPLYKYYEE TGNYLTKYSK KDNGPVIKKI KYYGNKLNAH LDITDDYPNS

901 RNKWKLSLK PYRFDVYLDN GVYKFVTVKN LDVIKKENYY EVNSKCYEEA KKLKKISNQA

961 EFIASFYNND LIKINGELYR VIGVNNDLLN RIEV MIDIT YREYLENMND KRPPRIIKTI1021

ASKTQSIKKY STDILGNLYE VKSKKHPQII KKG (SEQ ID NO: 14497).

[067] In certain embodiments, the Cas9 is an inactivated Cas9 (dCas9). In certain embodiments, the inactivated Cas9 (dCas9) is isolated or derived from Staphylococcus pyogenes and comprises DIOA and H840A within the catalytic site. In certain embodiments, the dCas9 comprises the amino acid sequence of:

1 XDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61 ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121 NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181 VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241 LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301 LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361 GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421 AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481 WDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL 541 SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601 IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661 RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL

721 HEHIANLAGS PAIKKGILQT VKWDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781 MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841 IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEWKKMK NYWRQLLNAK LITQRKFDNL

901 TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961 KLVSDFRKDF QFYKVREINN YHHAHDAYLN AWGTALIKK YPKLESEFVY GDYKVYDVRK

1021 MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081 ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141 YSVLWAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201 YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE

1261 QHKHYLDEI I EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA

1321 PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD (SEQ ID NO:

14498) .

[068] In certain embodiments, the Cas9 is an inactivated Cas9 (dCas9). In certain embodiments, the inactivated Cas9 (dCas9) is isolated or derived from Staphylococcus pyogenes and comprises DIOA and H840A within the catalytic site. In certain embodiments, the dCas9 comprises the amino acid sequence of:

1 MDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61 ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121 NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181 VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241 LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301 LLSDILRV T EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361 GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421 AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481 WDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541 SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601 IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661 RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL

721 HEHIANLAGS PAIKKGILQT VKWDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781 MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841 IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEWKKMK NYWRQLLNAK LITQRKFDNL

901 TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961 KLVSDFRKDF QFYKVREINN YHHAHDAYLN AWGTALIKK YPKLESEFVY GDYKVYDVRK

1021 MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081 ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141 YSVLWAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201 YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE 1261 QHKHYLDEI I EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA 1321 PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD (SEQ ID NO: 14499) .

[069] The disclosure provides an immune cell modified according to the method of the disclosure. The immune cell may be a T-lymphocyte, a Natural Killer (NK) cell, a Cytokine- induced Killer (CIK) cell or a Natural Killer T (NKT) cell. The immune cell may be further modified by a second gene editing tool, including, but not limited to those gene editing tools comprising an endonuclease operably-linked to either a Cas9 or a TALE sequence.

Alternatively or in addition, the second gene editing tool may include an excision-only piggyBac transposase to re-excise the inserted sequences or any portion thereof. For example, the excision-only piggyBac transposase may be used to "re-excise" the transposon.

[070] In certain embodiments, the transposon is a piggyBac transposon. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™ or a Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a Super piggyBac™ (SPB) transposase, the sequence encoding the transposase is an mRNA sequence.

[071] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme. The piggyBac (PB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG 61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG 121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF 181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV 241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD 301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ 361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC 421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN 481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO: 14487) .

[072] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at one or more of positions 30, 165, 282, or 538 of the sequence: 1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV

541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO

144ί 37) .

[073] In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at two or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at three or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at each of the following positions 30, 165, 282, and 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 165 of the sequence of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G). In certain embodiments, the amino acid substitution at position 282 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 538 of the sequence of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N).

[074] In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) transposase enzyme. In certain embodiments, the Super piggyBac™ (SPB) transposase enzymes of the disclosure may comprise or consist of the amino acid sequence of the sequence of SEQ ID NO: 14487 wherein the amino acid substitution at position 30 is a substitution of a valine (V) for an isoleucine (I), the amino acid substitution at position 165 is a substitution of a serine (S) for a glycine (G), the amino acid substitution at position 282 is a substitution of a valine (V) for a methionine (M), and the amino acid substitution at position 538 is a substitution of a lysine (K) for an asparagine (N). In certain embodiments, the Super piggyBac™ (SPB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG 61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG 121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF 181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV 241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD 301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ 361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC 421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN 481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO: 14484) .

[075] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 3, 46, 82, 103, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 258, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 486, 503, 552, 570 and 591 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 46, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 485, 503, 552 and 570. In certain embodiments, the amino acid substitution at position 3 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for a serine (S). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an alanine (A). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 82 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 119 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for an arginine (R). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) a cysteine (C). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a

phenylalanine (F). In certain embodiments, the amino acid substitution at position 185 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 187 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for an alanine (A). In certain embodiments, the amino acid substitution at position 200 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a phenylalanine (F).In certain embodiments, the amino acid substitution at position 207 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a valine (V). In certain embodiments, the amino acid substitution at position 209 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a valine (V). In certain embodiments, the amino acid substitution at position 226 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a methionine (M). In certain embodiments, the amino acid substitution at position 235 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a leucine (L). In certain embodiments, the amino acid substitution at position 240 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 241 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 243 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a proline (P). In certain embodiments, the amino acid substitution at position 258 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a proline (P). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine for a proline (P). In certain embodiments, the amino acid substitution at position 315 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for an arginine (R).In certain embodiments, the amino acid substitution at position 319 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a threonine (T). In certain embodiments, the amino acid substitution at position 327 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 328 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a cysteine (C). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 421 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for the aspartic acid (D). In certain embodiments, the amino acid substitution at position 436 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a valine (V). In certain embodiments, the amino acid substitution at position 456 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a methionine (M). In certain embodiments, the amino acid substitution at position 470 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 485 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a serine (S). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a methionine (M). In certain embodiments, the amino acid substitution at position 552 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a glutamine (Q). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a glutamine (Q).

In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at two, three, four, five, six or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 194 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 372 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for an arginine (R). In certain embodiments, the amino acid substitution at position 375 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a lysine (K). In certain embodiments, the amino acid substitution at position 450 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for an aspartic acid (D). In certain embodiments, the amino acid substitution at position 509 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a serine (S). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487. In certain embodiments, including those embodiments wherein the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, the piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 372, 375 and 450 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, and a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487. In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487 and a substitution of an asparagine (N) for an aspartic acid (D) at position 450 of SEQ ID NO: 14487.

[076] The disclosure provides a culture media for enhancing viability of a modified immune cell comprising IL-2, IL-21, IL-7, IL-15 or any combination thereof. The modified immune cell may be a T-lymphocyte, a Natural Killer (NK) cell, a Cytokine-induced Killer (CIK) cell or a Natural Killer T (NKT) cell. In some embodiments, the modified immune cell is a T- lymphocyte. In some embodiments, the T-lymphocyte is an early memory T-cell. In some embodiments, the T-lymphocyte is a stem cell-like T-cell. In some embodiments, the T- lymphocyte is a stem memory T cell (TSCM). In some embodiments, the T-lymphocyte is a central memory T cell (TCM). The modified immune cell may contain one or more exogenous DNA sequences. The modified immune cell may contain one or more exogenous RNA sequences. The modified immune cell may have been electroporated or nucleofected.

BRIEF DESCRIPTION OF THE DRAWINGS [077] Figure 1 is a series of graphs depicting transfection efficiency and cell viability following plasmid DNA nucleofection in primary human T lymphocytes.

[078] Figure 2 is a series of graphs depicting DNA cytotoxicity to T cells.

[079] Figure 3 is a series of graphs showing that DNA-mediated cytotoxicity in T cells is dose dependent.

[080] Figure 4 is a series of graphs showing that extracellular plasmid DNA is not cytotoxic.

[081] Figure 5 is a series of graphs depicting efficient transposition using SPB mRNA in Jurkat cells.

[082] Figure 6 is a series of graphs depicting efficient transposition in T lymphocytes using SPB mRNA.

[083] Figure 7 is a series of graphs depicting efficient delivery of linearized DNA transposon products.

[084] Figure 8 is a series of graphs showing that addition of that IL-7 and IL-15 and immediate stimulation of T cells post-nucleofection enhances cell viability.

[085] Figure 9 is a series of graphs showing that IL-7 and IL-15 rescue T cells from DNA mediated toxicity

[086] Figure 10 is a series of graphs showing that immediate stimulation of T cells post- nucleofection enhances cell viability.

[087] Figure 1 1 A-C is a series of graphs depicting T cell transposition with varying amounts of DNA. Primary human pan T cells were nucleofected with varying amounts of DNA using piggyBac™. T cells were nucleofected with the indicated amounts of transposon and 5 μg SPB mRNA. Cells were then stimulated on day 2 post-nucleofection through CD3 and CD28. As expected, T cells nucleofected with high amounts of DNA exhibited high episomal expression at day 1 post nucleofection whereas almost no episomal expression was observed at low DNA doses. In contrast, following expansion at day 21 post nucleofection the greatest percentage of transgene positive cells were observed in lower DNA amounts peaking at 1.67 μg for this transposon. (A) Flow analysis for transgene positive cells at day 1 and 21. (B) Percentage of transgene positive T cells. (C) Percentage of viable T cells at day 1 and 21. For all graphs shown in this figure, the Y-axis ranges from 0 to 100% in increments of 20% and the X-axis ranges from 0 to 10 ⁵ by powers of 10.

[088] Figure 12A-B is a series of graphs depicting T cell transposition with low DNA amounts using the Sleeping Beauty™ 100X (SB100X) transposase. Primary human pan T cells were nucleofected with GFP plasmids encoding either the piggyBac™ (PB) or Sleeping Beauty™ (SB) ITRs. (A) Cells were nucleofected with the indicated amounts of SB transposon and 1 μg SB transposase mRNA. (B) Cells were nucleofected with the indicated amounts of SB transposase and 0.75 μg SB transposon. Flow analysis was performed on day 14 post nucleofection for all samples. For all graphs shown in this figure, the Y-axis ranges from 0 to 250K in increments of 50K and the X-axis ranges from 0 to 10 ⁵ by powers of 10.

[089] Figure 13A is a series of plots depicting T cells transposed with a plasmid containing a sequence encoding a transposon comprising a sequence encoding an inducible caspase polypeptide (a safety switch, "iC9"), a CARTyrin (anti-BCMA), and a selectable marker. Left-hand plots depict live T cells exposed to transposase in the absence of the plasmid. Right-hand plots depict live T cells exposed to transposase in the presence of the plasmid. Cells were exposed to either a hyperactive transposase (the "Super piggyBac") or a wild type piggyBac transposase.

[090] Figure 13B is a series of plots depicting T cells transposed with a plasmid containing a sequence encoding a green fluorescent protein (GFP). Left-hand plots depict live T cells exposed to transposase in the absence of the plasmid. Right-hand plots depict live T cells exposed to transposase in the presence of the plasmid. Cells were exposed to either a hyperactive transposase (the "Super piggyBac") or a wild type piggyBac transposase.

[091] Figure 13C is a table depicting the percent of transformed T cells resulting from transposition with WT versus hyperactive piggyBac transposase. T cells contacted with the hyperactive piggyBac transposase (the Super piggyBac transposase) were transformed at a rate 4-fold greater than WT transposase.

[092] Figure 13D is a graph depicting the percent of transformed T cells resulting from transposition with WT versus hyperactive piggyBac transposase 5 days after nucleofection. T cells contacted with the hyperactive piggyBac transposase (the Super piggyBac transposase) were transformed at a rate far greater than WT transposase.

[093] Figure 14 is a graph depicting transposition in natural killer (NK) cells. Transposition of non-activated NK cells derived from CD3 -depleted leukopheresis (containing

CD14/CD19/CD56+ cells) is shown. Cells were electroporated (EP) with plasmid piggyBac transposon DNA encoding GFP and mRNA encoding super piggyBac. The program from Lonza 4D nucleofector or BTX ECM 830 (500V, 700 usee pulse length, 0.2 mm electrode gap, one pulse) is indicated on the X-axis. Transposed cells were co-cultured (stimulated) at day 2 with artificial antigen presenting cells (aAPCs). Fluorescent activated cell sorting (FACS) analysis of percent GFP positive cells at day 7 post-EP (day 5 post-stim) is indicated on the Y-axis with gray bars. Percent viability as shown by percent 7-Aminoactinomycin D (7AAD)-negative cells at day 2 post-EP is indicated on the Y-axis with gray bars.

[094] Figure 15A-B are a series of 10 FACs plots (Figure 15 A) and a graph (Figure 15B) showing transposon titration for transposition in natural killer (NK) cells. Transposition of non-activated NK cells from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells) is shown. Cells were electroporated with a plasmid piggyBac transposon encoding GFP at amounts ranging from 0 to 10 ug of DNA and 5 ug mRNA encoding Super piggyBac using the indicated Maxcyte electroporator program. Transposed cells were stimulated at day 2 with artificial antigen presenting cells (aAPCs). Figure 15A FACs plots top row shows CD56+ (y-axis) versus GFP+ (x-axis) expression, while the bottom row shows 7AAD (y- axis) versus forward scatter (FSC, x-axis). Figure 15B is a bar graph analysis of the percentage of GFP+ cells of CD56+ cells at day 6 post-electroporation (EP) and day 4 post- stimulation (black bars), and the percent viability as shown by 7AAD-negative cells at day 2 post EP (gray bars).

[095] Figure 16A-B are a series of 7 FACs plots (Figure 16A) and a graph (Figure 16B) showing dose-dependent DNA-mediated cytotoxicity in NK cells. FACS analysis of live cells (7AAD-negative/FSC) at day 2 post-EP using the Lonza 4D Nucleofector program DN- 100 are shown (Figure 16A). FACS plots (Figure 16A) are quantified in a graph (Figure 16B). 5E6 cells per EP were electroporated in 100 uL P3 buffer in cuvettes. Cells were electroporated with no DNA (Mock) or varying amounts of piggyBac GFP transposon co- delivered with 5 ug Super piggyBac mRNA.

[096] Figure 17 is a series of 5 graphs showing the in vitro differentiation of piggyBac modified hematopoietic stem and precursor cells (HSPCs) into B cells. Human CD34+ HSPCs were electroporated with mRNA encoding Super piggyBac along with a piggyBac transposon encoding GFP. After electroporation, HSPCs were primed for B cell

differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for 5 days. On day 6, cells were transferred to a layer of MS-5 feeder cells and fed bi-weekly, along with transfer to a fresh layer of feeders once per week. On day 34 of the in vitro differentiation process, CD 19+ B cells were generated and detectable in the culture. Top row: FACs plots showing CD 19 (y-axis) and CD34 (x-axis) in, from left to right, human primary bone marrow cells, at day 6 of in vitro differentiation, and at day 34 of in vitro differentiation. Bottom row: graphs depicting GFP expression in the indicated boxed populations of cells from the FACs plots in the top row at days 6 and 34 of in vitro differentiation. [097] Figure 18 is a schematic depiction of the Csy4-T2A-Clo051 -G4Slinker-dCas9 construct map.

[098] Figure 19 is a schematic depiction of the pRTl-Clo051-dCas9 Double NLS construct map.

DETAILED DESCRIPTION

[099] Disclosed are compositions and methods for the ex-vivo genetic modification of an immune cell or a precursor thereof comprising delivering to the immune cell or immune precursor cell, (a) a nucleic acid or amino acid sequence comprising a sequence encoding a transposase enzyme and (b) a recombinant and non-naturally occurring DNA sequence comprising a DNA sequence encoding a transposon. In certain embodiments, the method further comprises the step of stimulating the immune cell or immune precursor cell with one or more cytokine(s).

Immune and Immune Precursor Cells

[0100] In certain embodiments, immune cells of the disclosure comprise lymphoid progenitor cells, natural killer (NK) cells, T lymphocytes (T-cell), stem memory T cells (TSCM cells), Stem cell-like T cells, B lymphocytes (B-cells), myeloid progenitor cells, neutrophils, basophils, eosinophils, monocytes, macrophages, platelets, erythrocytes, red blood cells (RBCs), megakaryocytes or osteoclasts.

[0101] In certain embodiments, immune precursor cells comprise any cells which can differentiate into one or more types of immune cells. In certain embodiments, immune precursor cells comprise multipotent stem cells that can self renew and develop into immune cells. In certain embodiments, immune precursor cells comprise hematopoietic stem cells (HSCs) or descendants thereof. In certain embodiments, immune precursor cells comprise precursor cells that can develop into immune cells. In certain embodiments, the immune precursor cells comprise hematopoietic progenitor cells (HPCs).

Hematopoietic Stem Cells (HSCs)

[0102] Hematopoietic stem cells (HSCs) are multipotent, self-renewing cells. All differentiated blood cells from the lymphoid and myeloid lineages arise from HSCs. HSCs can be found in adult bone marrow, peripheral blood, mobilized peripheral blood, peritoneal dialysis effluent and umbilical cord blood.

[0103] HSCs of the disclosure may be isolated or derived from a primary or cultured stem cell. HSCs of the disclosure may be isolated or derived from an embryonic stem cell, a multipotent stem cell, a pluripotent stem cell, an adult stem cell, or an induced pluripotent stem cell (iPSC).

[0104] Immune precursor cells of the disclosure may comprise an HSC or an HSC descendent cell. Exemplary HSC descendent cells of the disclosure include, but are not limited to, multipotent stem cells, lymphoid progenitor cells, natural killer (NK) cells, T lymphocyte cells (T-cells), B lymphocyte cells (B-cells), myeloid progenitor cells, neutrophils, basophils, eosinophils, monocytes, and macrophages.

[0105] HSCs produced by the methods of the disclosure may retain features of "primitive" stem cells that, while isolated or derived from an adult stem cell and while committed to a single lineage, share characteristics of embryonic stem cells. For example, the "primitive" HSCs produced by the methods of the disclosure retain their "sternness" following division and do not differentiate. Consequently, as an adoptive cell therapy, the "primitive" HSCs produced by the methods of the disclosure not only replenish their numbers, but expand in vivo. "Primitive" HSCs produced by the methods of the disclosure may be therapeutically - effective when administered as a single dose. In some embodiments, primitive HSCs of the disclosure are CD34+. In some embodiments, primitive HSCs of the disclosure are CD34+ and CD38-. In some embodiments, primitive HSCs of the disclosure are CD34+, CD38- and CD90+. In some embodiments, primitive HSCs of the disclosure are CD34+, CD38-, CD90+ and CD45RA-. In some embodiments, primitive HSCs of the disclosure are CD34+, CD38-, CD90+, CD45RA-, and CD49f+. In some embodiments, the most primitive HSCs of the disclosure are CD34+, CD38-, CD90+, CD45RA-, and CD49f+.

[0106] In some embodiments of the disclosure, primitive HSCs, HSCs, and/or HSC descendent cells may be modified according to the methods of the disclosure to express an exogenous sequence (e.g. a chimeric antigen receptor or therapeutic protein). In some embodiments of the disclosure, modified primitive HSCs, modified HSCs, and/or modified HSC descendent cells may be forward differentiated to produce a modified immune cell including, but not limited to, a modified T cell, a modified natural killer cell and/or a modified B-cell of the disclosure.

T Cells

[0107] Modified T cells of the disclosure may be derived from modified hematopoietic stem and progenitor cells (HSPCs) or modified HSCs.

[0108] Unlike traditional biologies and chemotherapeutics, modified-T cells of the disclosure possess the capacity to rapidly reproduce upon antigen recognition, thereby potentially obviating the need for repeat treatments. To achieve this, in some embodiments, modified-T cells of the disclosure not only drive an initial response, but also persist in the patient as a stable population of viable memory T cells to prevent potential relapses.

Alternatively, in some embodiments, when it is not desired, modified-T cells of the disclosure do not persist in the patient.

[0109] Intensive efforts have been focused on the development of antigen receptor molecules that do not cause T cell exhaustion through antigen-independent (tonic) signaling, as well as of a modified-T cell product containing early memory T cells, especially stem cell memory (TSCM) or stem cell-like T cells. Stem cell-like modified-T cells of the disclosure exhibit the greatest capacity for self-renewal and multipotent capacity to derive central memory (TCM) T cells or TCM like cells, effector memory (TEM) and effector T cells (TE), thereby producing better tumor eradication and long-term modified-T cell engraftment. A linear pathway of differentiation may be responsible for generating these cells: Naive T cells (TN) > TSCM > TCM > TEM > TE > TTE, whereby TN is the parent precursor cell that directly gives rise to TSCM, which then, in turn, directly gives rise to TCM, etc. Compositions of T cells of the disclosure may comprise one or more of each parental T cell subset with TSCM cells being the most abundant (e.g. TSCM > TCM > TEM > TE > TTE).

[0110] In some embodiments of the methods of the disclosure, the immune cell precursor is differentiated into or is capable of differentiating into an early memory T cell, a stem cell like T-cell, a Naive T cells (TN), a TSCM, a TCM, a TEM, a TE, or a TTE. In some embodiments, the immune cell precursor is a primitive HSC, an HSC, or a HSC descendent cell of the disclosure.

[0111] In some embodiments of the methods of the disclosure, the immune cell is an early memory T cell, a stem cell like T-cell, a Naive T cells (TN), a TSCM, a TCM, a TEM, a TE, or a TTE.

[0112] In some embodiments of the methods of the disclosure, the immune cell is an early memory T cell.

[0113] In some embodiments of the methods of the disclosure, the immune cell is a stem cell like T-cell.

[0114] In some embodiments of the methods of the disclosure, the immune cell is a TSCM.

[0115] In some embodiments of the methods of the disclosure, the immune cell is a TCM.

[0116] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of an early memory T cell. In certain embodiments, the plurality of modified early memory T cells comprises at least one modified stem cell-like T cell. In certain embodiments, the plurality of modified early memory T cells comprises at least one modified TSCM. In certain embodiments, the plurality of modified early memory T cells comprises at least one modified TCM.

[0117] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of a stem cell-like T cell. In certain embodiments, the plurality of modified stem cell-like T cells comprises at least one modified TSCM. In certain embodiments, the plurality of modified stem cell-like T cells comprises at least one modified TCM.

[0118] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of a stem memory T cell (TSCM). In certain embodiments, the cell-surface markers comprise CD62L and CD45RA. In certain embodiments, the cell-surface markers comprise one or more of CD62L, CD45RA, CD28, CCR7, CD127, CD45RO, CD95, CD95 and IL-2R{$. In certain embodiments, the cell-surface markers comprise one or more of CD45RA, CD95, IL-2R , CCR7, and CD62L.

[0119] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of a central memory T cell (TCM). In certain embodiments, the cell-surface markers comprise one or more of CD45RO, CD95, IL-2R , CCR7, and CD62L.

[0120] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of a naive T cell (TN). In certain embodiments, the cell-surface markers comprise one or more of CD45RA, CCR7 and CD62L. [0121] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of an effector T-cell (modified TEFF). In certain embodiments, the cell- surface markers comprise one or more of CD45RA, CD95, and IL-2R{$.

[0122] In some embodiments of the methods of the disclosure, the methods modify and/or the methods produce a plurality of modified T cells, wherein at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of modified T cells expresses one or more cell- surface marker(s) of a stem cell-like T cell, a stem memory T cell (TSCM) or a central memory T cell (TCM).

[0123] In some embodiments of the methods of the disclosure, a buffer comprises the immune cell or precursor thereof. The buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the immune cell or precursor thereof, including T-cells. In certain embodiments, the buffer maintains or enhances a level of cell viability and/or a stemlike phenotype of the primary human T cells prior to the nucleofection. In certain embodiments, the buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the primary human T cells during the nucleofection. In certain embodiments, the buffer maintains or enhances a level of cell viability and/or a stem-like phenotype of the primary human T cells following the nucleofection. In certain embodiments, the buffer comprises one or more of KCl, MgCh, ClNa, Glucose and Ca(N03)2 in any absolute or relative abundance or concentration, and, optionally, the buffer further comprises a supplement selected from the group consisting of HEPES, Tris/HCl, and a phosphate buffer. In certain embodiments, the buffer comprises 5 mM KCl, 15 mM MgCh, 90 mM ClNa, 10 mM Glucose and 0.4 mM Ca(NCb)2. In certain embodiments, the buffer comprises 5 mM KCl, 15 mM MgCh, 90 mM ClNa, 10 mM Glucose and 0.4 mM Ca(N0 ₃ )2 and a supplement comprising 20 mM HEPES and 75 mM Tris/HCl. In certain embodiments, the buffer comprises 5 mM KCl, 15 mM MgCh, 90 mM ClNa, 10 mM Glucose and 0.4 mM Ca(N0 ₃ )2 and a supplement comprising 40 mM Na2HP04/NaH2P04 at pH 7.2. In certain embodiments, the composition comprising primary human T cells comprises 100 μΐ of the buffer and between 5xl0 ⁶ and 25xl0 ⁶ cells. In certain embodiments, the composition comprises a scalable ratio of 250e6 primary human T cells per milliliter of buffer or other media during the introduction step. [0124] In some embodiments of the methods of the disclosure, the introducing step may comprise delivery of transposon and/or transposase by a method other than electroporation or nucleofection. In some embodiments, a composition comprises a scalable ratio of 250e6 primary human T cells per milliliter of buffer or other media during the introduction step.

[0125] In some embodiments of the methods of the disclosure, the introducing step comprises one or more of topical delivery, adsorption, absorption, electroporation, spin- fection, co-culture, transfection, mechanical delivery, sonic delivery, vibrational delivery, magnetofection or by nanoparticle-mediated delivery.

[0126] In some embodiments of the methods of the disclosure, the introducing step comprises liposomal transfection, calcium phosphate transfection, fugene transfection, and dendrimer-mediated transfection.

[0127] In some embodiments of the methods of the disclosure, the introducing step comprises mechanical transfection comprises cell squeezing, cell bombardment, or gene gun techniques.

[0128] In some embodiments of the methods of the disclosure, the introducing step comprises nanoparticle-mediated transfection comprises liposomal delivery, delivery by micelles, and delivery by polymerosomes.

[0129] In some embodiments of the methods of the disclosure, the methods comprise contacting an immune cell of the disclosure, including a T cell of the disclosure, and a T-cell expansion composition. In some embodiments of the methods of the disclosure, the step of introducing a transposon and/or transposase of the disclosure into an immune cell of the disclosure may further comprise contacting the immune cell and a T-cell expansion composition. In some embodiments, including those in which the introducing step of the methods comprises an electroporation or a nucleofection step, the electroporation or a nucleofection step may be performed with the immune cell contacting T-cell expansion composition of the disclosure.

[0130] In some embodiments of the methods of the disclosure, the T-cell expansion composition comprises, consists essentially of or consists of phosphorus; one or more of an octanoic acid, a palmitic acid, a linoleic acid, and an oleic acid; a sterol; and an alkane.

[0131] In certain embodiments of the methods of producing a modified T cell of the disclosure, the expansion supplement comprises one or more cytokine(s). The one or more cytokine(s) may comprise any cytokine, including but not limited to, lymphokines.

Exemplary lympokines include, but are not limited to, interleukin-2 (IL-2), interleukin-3 (IL- 3), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-7 (IL-7), interleukin-15 (IL-15), interleukin-21 (IL-21), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (INFy). The one or more cytokine(s) may comprise IL-2.

[0132] In some embodiments of the methods of the disclosure, the T-cell expansion composition comprises human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid, nicotinamide, 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD), diisopropyl adipate (DIP A), n- butyl-benzenesulfonamide, 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester, palmitic acid, linoleic acid, oleic acid, stearic acid hydrazide, oleamide, a sterol and an alkane. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid, palmitic acid, linoleic acid, oleic acid and a sterol. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of between 0.9 mg/kg to 90 mg/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; oleic acid at a concentration of 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; and a sterol at a concentration of about 0.1 mg/kg to 10 mg/kg, inclusive of the endpoints. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of about 9 mg/kg, palmitic acid at a concentration of about 2 mg/kg, linoleic acid at a concentration of about 2 mg/kg, oleic acid at a concentration of about 2 mg/kg and a sterol at a concentration of about 1 mg/kg. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of between 6.4 μηιοΐ/kg and 640 μηιοΐ/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.7 μηιοΐ/kg and 70 μηιοΐ/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.75 μηιοΐ/kg and 75 μηιοι/kg, inclusive of the endpoints; oleic acid at a concentration of between 0.75 μηιοΐ/kg and 75 μηιοΐ/kg, inclusive of the endpoints; and a sterol at a concentration of between 0.25 μηιοΐ/kg and 25 μηιοΐ/kg, inclusive of the endpoints. In certain embodiments of this method, the T-cell expansion composition further comprises one or more of octanoic acid at a concentration of about 64 μηιοΐ/kg, palmitic acid at a concentration of about 7 μηιοΐ/kg, linoleic acid at a concentration of about 7.5 μηιοΐ/kg, oleic acid at a concentration of about 7.5 μηιοι/kg and a sterol at a concentration of about 2.5 μηιοι/kg. [0133] In certain embodiments, the T-cell expansion composition comprises one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement to produce a plurality of expanded modified T-cells, wherein at least 2% of the plurality of modified T-cells expresses one or more cell-surface marker(s) of an early memory T cell, a stem cell-like T cell, a stem memory T cell (TSCM) and/or a central memory T cell (TCM). In certain embodiments, the T-cell expansion composition comprises or further comprises one or more of octanoic acid, nicotinamide, 2,4,7,9-tetramethyl-5-decyn- 4,7-diol (TMDD), diisopropyl adipate (DIP A), n-butyl-benzenesulfonamide, 1,2- benzenedicarboxylic acid, bis(2-methylpropyl) ester, palmitic acid, linoleic acid, oleic acid, stearic acid hydrazide, oleamide, a sterol and an alkane. In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid, palmitic acid, linoleic acid, oleic acid and a sterol (e.g. cholesterol). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of between 0.9 mg/kg to 90 mg/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; oleic acid at a concentration of 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; and a sterol at a concentration of about 0.1 mg/kg to 10 mg/kg, inclusive of the endpoints (wherein mg/kg = parts per million). In certain

embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of about 9 mg/kg, palmitic acid at a concentration of about 2 mg/kg, linoleic acid at a concentration of about 2 mg/kg, oleic acid at a concentration of about 2 mg/kg, and a sterol at a concentration of about 1 mg/kg (wherein mg/kg = parts per million). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of about 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of about 1.01 mg/kg (wherein mg/kg = parts per million). In certain embodiments, the T-cell expansion composition comprises octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of 1.01 mg/kg (wherein mg/kg = parts per million). In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of between 6.4 μιηοΐ/kg and 640 μιηοΐ/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.7 μιηοΐ/kg and 70 μιηοΐ/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.75 μιηοΐ/kg and 75 μιηοΐ/kg, inclusive of the endpoints; oleic acid at a concentration of between 0.75 μηιοΐ/kg and 75 μηιοΐ/kg, inclusive of the endpoints; and a sterol at a concentration of between 0.25 μηιοΐ/kg and 25 μηιοΐ/kg, inclusive of the endpoints. In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of about 64 μηιοΐ/kg, palmitic acid at a concentration of about 7 μηιοΐ/kg, linoleic acid at a concentration of about 7.5 μηιοΐ/kg, oleic acid at a concentration of about 7.5 μηιοΐ/kg and a sterol at a concentration of about 2.5 μηιοΐ/kg. In certain embodiments, the T-cell expansion composition comprises one or more of octanoic acid at a concentration of about 63.75 μηιοΐ/kg, palmitic acid at a concentration of about 7.27 μηιοΐ/kg, linoleic acid at a concentration of about 7.57 μηιοΐ/kg, oleic acid at a concentration of about 7.56 μηιοΐ/kg and a sterol at a concentration of about 2.61 μηιοΐ/kg. In certain embodiments, the T-cell expansion composition comprises octanoic acid at a concentration of about 63.75 μιηοΐ/kg, palmitic acid at a concentration of about 7.27 μηιοΐ/kg, linoleic acid at a concentration of about 7.57 μιηοΐ/kg, oleic acid at a concentration of 7.56 μιηοΐ/kg and a sterol at a concentration of 2.61 μηιοΐ/kg.

[0134] As used herein, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement at 37°C. Alternatively, or in addition, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used

interchangeably with a media comprising one or more of phosphorus, an octanoic fatty acid, a palmitic fatty acid, a linoleic fatty acid and an oleic acid. In certain embodiments, the media comprises an amount of phosphorus that is 10-fold higher than may be found in, for example, Iscove's Modified Dulbecco's Medium ((IMDM); available at ThermoFisher Scientific as Catalog number 12440053).

[0135] As used herein, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, Iscove's MDM, and an expansion supplement at 37°C. Alternatively, or in addition, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of the following elements: boron, sodium, magnesium, phosphorus, potassium, and calcium. In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of the following elements present in the corresponding average concentrations: boron at 3.7 mg/L, sodium at 3000 mg/L, magnesium at 18 mg/L, phosphorus at 29 mg/L, potassium at 15 mg/L and calcium at 4 mg/L.

[0136] As used herein, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement at 37°C. Alternatively, or in addition, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used

interchangeably with a media comprising one or more of the following components: octanoic acid (CAS No. 124-07-2), nicotinamide (CAS No. 98-92-0), 2,4,7,9-tetramethyl-5-decyn-4,7- diol (TMDD) (CAS No. 126-86-3), diisopropyl adipate (DIP A) (CAS No. 6938-94-9), n- butyl-benzenesulfonamide (CAS No. 3622-84-2), 1 ,2-benzenedicarboxylic acid, bis(2- methylpropyl) ester (CAS No. 84-69-5), palmitic acid (CAS No. 57-10-3), linoleic acid (CAS No. 60-33-3), oleic acid (CAS No. 112-80-1), stearic acid hydrazide (CAS No. 4130-54-5), oleamide (CAS No. 3322-62-1), sterol (e.g., cholesterol) (CAS No. 57-88-5), and alkanes (e.g., nonadecane) (CAS No. 629-92-5). In certain embodiments, the terms "supplemented T- cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of the following components: octanoic acid (CAS No. 124-07-2), nicotinamide (CAS No. 98-92-0), 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD) (CAS No. 126-86-3), diisopropyl adipate (DIP A) (CAS No. 6938-94-9), n-butyl- benzenesulfonamide (CAS No. 3622-84-2), 1 ,2-benzenedicarboxylic acid, bis(2- methylpropyl) ester (CAS No. 84-69-5), palmitic acid (CAS No. 57-10-3), linoleic acid (CAS No. 60-33-3), oleic acid (CAS No. 112-80-1), stearic acid hydrazide (CAS No. 4130-54-5), oleamide (CAS No. 3322-62-1), sterol (e.g., cholesterol) (CAS No. 57-88-5), alkanes (e.g., nonadecane) (CAS No. 629-92-5), and phenol red (CAS No. 143-74-8). In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of the following components: octanoic acid (CAS No. 124-07-2), nicotinamide (CAS No. 98-92-0), 2,4,7,9-tetramethyl-5-decyn-4,7-diol (TMDD) (CAS No. 126-86-3), diisopropyl adipate (DIP A) (CAS No. 6938-94-9), n-butyl-benzenesulfonamide (CAS No. 3622-84-2), 1 ,2- benzenedicarboxylic acid, bis(2-methylpropyl) ester (CAS No. 84-69-5), palmitic acid (CAS No. 57-10-3), linoleic acid (CAS No. 60-33-3), oleic acid (CAS No. 112-80-1), stearic acid hydrazide (CAS No. 4130-54-5), oleamide (CAS No. 3322-62-1), phenol red (CAS No. 143- 74-8) and lanolin alcohol. [0137] In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2- Mercaptoethanol, and an expansion supplement at 37°C. Alternatively, or in addition, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of the following ions: sodium, ammonium, potassium, magnesium, calcium, chloride, sulfate and phosphate.

[0138] As used herein, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, and an expansion supplement at 37°C. Alternatively, or in addition, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used

interchangeably with a media comprising one or more of the following free amino acids: histidine, asparagine, serine, glutamate, arginine, glycine, aspartic acid, glutamic acid, threonine, alanine, proline, cysteine, lysine, tyrosine, methionine, valine, isoleucine, leucine, phenylalanine and tryptophan. In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of the following free amino acids in the corresponding average mole percentages: histidine (about 1%), asparagine (about 0.5%), serine (about 1.5%), glutamine (about 67%), arginine (about 1.5%), glycine (about 1.5%), aspartic acid (about 1 %), glutamic acid (about 2%), threonine (about 2%), alanine (about 1 %), proline (about 1.5%), cysteine (about 1.5%), lysine (about 3%), tyrosine (about 1.5%), methionine (about 1 %), valine (about 3.5%), isoleucine (about 3%), leucine (about 3.5%), phenylalanine (about 1.5%) and tryptophan (about 0.5%). In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used

interchangeably with a media comprising one or more of the following free amino acids in the corresponding average mole percentages: histidine (about .78%), asparagine (about 0.4%), serine (about 1.6%), glutamine (about 67.01%), arginine (about 1.67%), glycine (about 1.72%), aspartic acid (about 1.00%), glutamic acid (about 1.93%), threonine (about 2.38%), alanine (about 1.11 %), proline (about 1.49%), cysteine (about 1.65%), lysine (about 2.84%), tyrosine (about 1.62%), methionine (about 0.85%), valine (about 3.45%), isoleucine (about 3.14%), leucine (about 3.3%), phenylalanine (about 1.64%) and tryptophan (about 0.37%). [0139] As used herein, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of human serum albumin, recombinant human insulin, human transferrin, 2-Mercaptoethanol, Iscove's MDM, and an expansion supplement at 37°C. Alternatively, or in addition, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of phosphorus, an octanoic fatty acid, a palmitic fatty acid, a linoleic fatty acid and an oleic acid. In certain embodiments, the media comprises an amount of phosphorus that is 10-fold higher than may be found in, for example, Iscove's Modified Dulbecco's Medium ((IMDM); available at ThermoFisher Scientific as Catalog number 12440053).

[0140] In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid, palmitic acid, linoleic acid, oleic acid and a sterol (e.g. cholesterol). In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of between 0.9 mg/kg to 90 mg/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; oleic acid at a concentration of 0.2 mg/kg to 20 mg/kg, inclusive of the endpoints; and a sterol at a concentration of about 0.1 mg/kg to 10 mg/kg, inclusive of the endpoints (wherein mg/kg = parts per million). In certain embodiments, the terms

"supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a

concentration of about 9 mg/kg, palmitic acid at a concentration of about 2 mg/kg, linoleic acid at a concentration of about 2 mg/kg, oleic acid at a concentration of about 2 mg/kg, and a sterol at a concentration of about 1 mg/kg (wherein mg/kg = parts per million). In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of about 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of about 1.01 mg/kg (wherein mg/kg = parts per million). In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of 9.19 mg/kg, palmitic acid at a concentration of 1.86 mg/kg, linoleic acid at a concentration of 2.12 mg/kg, oleic acid at a concentration of about 2.13 mg/kg, and a sterol at a concentration of 1.01 mg/kg (wherein mg/kg = parts per million). In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of between 6.4 μηιοΐ/kg and 640 μηιοΐ/kg, inclusive of the endpoints; palmitic acid at a concentration of between 0.7 μηιοΐ/kg and 70 μηιοΐ/kg, inclusive of the endpoints; linoleic acid at a concentration of between 0.75 μηιοΐ/kg and 75 μηιοΐ/kg, inclusive of the endpoints; oleic acid at a concentration of between 0.75 μηιοΐ/kg and 75 μηιοΐ/kg, inclusive of the endpoints; and a sterol at a concentration of between 0.25 μηιοΐ/kg and 25 μηιοΐ/kg, inclusive of the endpoints. In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a

concentration of about 64 μηιοΐ/kg, palmitic acid at a concentration of about 7 μηιοΐ/kg, linoleic acid at a concentration of about 7.5 μηιοΐ/kg, oleic acid at a concentration of about 7.5 μιηοΐ/kg and a sterol at a concentration of about 2.5 μιηοι/kg.

[0141] In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of about 63.75 μηιοΐ/kg, palmitic acid at a concentration of about 7.27 μηιοΐ/kg, linoleic acid at a concentration of about 7.57 μηιοΐ/kg, oleic acid at a concentration of about 7.56 μηιοΐ/kg and a sterol at a concentration of about 2.61 μηιοΐ/kg. In certain embodiments, the terms "supplemented T-cell expansion composition" or "T-cell expansion composition" may be used interchangeably with a media comprising one or more of octanoic acid at a concentration of about 63.75 μηιοΐ/kg, palmitic acid at a concentration of about 7.27 μηιοΐ/kg, linoleic acid at a concentration of about 7.57 μηιοΐ/kg, oleic acid at a concentration of 7.56 μιηοΐ/kg and a sterol at a concentration of 2.61 μηιοΐ/kg.

[0142] Modified T-cells of the disclosure, including modified stem cell-like T cells, TSCM and/or TCM of the disclosure, may be incubated, cultured, grown, stored, or otherwise, combined at any step in the methods of the procedure with a growth medium comprising one or more inhibitors a component of a PI3K pathway. Exemplary inhibitors a component of a PI3K pathway include, but are not limited to, an inhibitor of GSK3 such as TWS 119 (also known as GSK 3B inhibitor XII; CAS Number 601514-19-6 having a chemical formula C18H14N4O2). Exemplary inhibitors of a component of a PI3K pathway include, but are not limited to, bb007 (BLUEBIRDBIO™). [0143] In some embodiments of the methods of the disclosure, the methods comprise contacting an immune cell of the disclosure and a T-cell activator composition. In some embodiments of the methods of the disclosure, the methods comprise contacting an immune cell precursor of the disclosure and a T-cell activator composition. In some embodiments of the methods of the disclosure, the methods comprise contacting a modified T cell of the disclosure and a T-cell activator composition. In some embodiments, the T-cell activator composition comprises one or more of an anti-human CD3 monospecific tetrameric antibody complex, an anti-human CD28 monospecific tetrameric antibody complex and an activation supplement to produce an activated modified T-cell or a plurality of activated modified T- cells. In some embodiments, the activated modified T-cell expresses one or more cell-surface marker(s) of an early memory T cell, a stem cell-like T cell, a TSCM or a TCM. In some embodiments, at least 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between of the plurality of activated modified T-cells express one or more cell-surface marker(s) of an early memory T cell, a stem cell-like T cell, a TSCM or a TCM.

[0144] In certain embodiments of the methods of producing a modified T cell (e.g. a stem cell-like T cell, a TSCM and/or a TCM) of the disclosure, the activation supplement may comprise one or more cytokine(s). The one or more cytokine(s) may comprise any cytokine, including but not limited to, lymphokines. Exemplary lympokines include, but are not limited to, interleukin-2 (IL-2), interleukin-3 (IL-3), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), interleukin-7 (IL-7), interleukin-15 (IL-15), interleukin-21 (IL-21), granulocyte-macrophage colony-stimulating factor (GM-CSF) and interferon-gamma (INFy). The one or more cytokine(s) may comprise IL-2.

Natural Killer (NK) cells

[0145] In certain embodiments, the modified immune or immune precursor cells of the disclosure are natural killer (NK) cells. In certain embodiments, NK cells are cytotoxic lymphocytes that differentiate from lymphoid progenitor cells.

[0146] Modified NK cells of the disclosure may be derived from modified hematopoietic stem and progenitor cells (HSPCs) or modified HSCs.

[0147]

[0148] In certain embodiments, non-activated NK cells are derived from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells).

[0149] In certain embodiments, NK cells are electroporated using a Lonza 4D nucleofector or BTX ECM 830 (500V, 700 usee pulse length, 0.2 mm electrode gap, one pulse). All Lonza 4D nucleofector programs are contemplated as within the scope of the methods of the disclosure.

[0150] In certain embodiments, 5x10E6 cells were electroporated per electroporation in 100 P3 buffer in cuvettes. However, this ratio of cells per volume is scalable for commercial manufacturing methods.

[0151] In certain embodiments, NK cells were stimulated by co-culture with an additional cell line. In certain embodiments, the additional cell line comprises artificial antigen presenting cells (aAPCs). In certain embodiments, stimulation occurs at day 1, 2, 3, 4, 5, 6, or 7 following electroporation. In certain embodiments, stimulation occurs at day 2 following electroporation.

[0152] In certain embodiments, NK cells express CD56.

B cells

[0153] In certain embodiments, the modified immune or immune precursor cells of the disclosure are B cells. B cells are a type of lymphocyte that express B cell receptors on the cell surface. B cell receptors bind to specific antigens.

[0154] Modified B cells of the disclosure may be derived from modified hematopoietic stem and progenitor cells (HSPCs) or modified HSCs.

[0155] In certain embodiments, HSPCs are modified using the methods of the disclosure, and then primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for at least 3 days, at least 4 days, at least 5 days, at least 6 days or at least 7 days. In certain embodiments, HSPCs are modified using the methods of the disclosure, and then primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for 5 days.

[0156] In certain embodiments, following priming, modified HSPC cells are transferred to a layer of feeder cells and fed bi-weekly, along with transfer to a fresh layer of feeders once per week. In certain embodiments, the feeder cells are MS -5 feeder cells.

[0157] In certain embodiments, modified HSPC cells are cultured with MS-5 feeder cells for at least 7, 14, 21, 28, 30, 33, 35, 42 or 48 days. In certain embodiments, modified HSPC cells were cultured with MS-5 feeder cells for 33 days.

Chimeric Antigen Receptors

[0158] In certain embodiments, a modified immune or pre-immune cell of the disclosure comprises a chimeric antigen receptor.

[0159] In certain embodiments of the methods of the disclosure, the recombinant and non- naturally occurring DNA sequence encoding a transposon further comprises a sequence encoding a chimeric antigen receptor or a portion thereof. Chimeric antigen receptors (CARs) of the disclosure may comprise (a) an ectodomain comprising an antigen recognition region, (b) a transmembrane domain, and (c) an endodomain comprising at least one costimulatory domain. In certain embodiments, the ectodomain may further comprise a signal peptide. Alternatively, or in addition, in certain embodiments, the ectodomain may further comprise a hinge between the antigen recognition region and the transmembrane domain. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD2, CD35, CD3s, CD3y, CO3C,, CD4, CD8a, CD19, CD28, 4-1BB or GM-CSFR signal peptide. In certain embodiments of the CARs of the disclosure, the signal peptide may comprise a sequence encoding a human CD8a signal peptide. In certain embodiments, the transmembrane domain may comprise a sequence encoding a human CD2, CD35, CD3s, CD3y, CO3C,, CD4, CD8a, CD19, CD28, 4-1BB or GM-CSFR transmembrane domain. In certain embodiments of the CARs of the disclosure, the transmembrane domain may comprise a sequence encoding a human CD8a transmembrane domain. In certain embodiments of the CARs of the disclosure, the endodomain may comprise a human CD3ζ endodomain.

[0160] In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a human 4-1BB, CD28, CD40, ICOS, MyD88, OX-40 intracellular segment, or any combination thereof. In certain embodiments of the CARs of the disclosure, the at least one costimulatory domain may comprise a CD28 and/or a 4-1BB costimulatory domain. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8a, IgG4, and/or CD4 sequence. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8a sequence.

[0161] The CD28 costimulatory domain may comprise an amino acid sequence comprising RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQE GLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPP

R (SEQ ID NO: 14659) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

RVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQE GLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPP

R (SEQ ID NO: 14659). The CD28 costimulatory domain may be encoded by the nucleic acid sequence comprising

cgcgtgaagtttagtcgatcagcagatgccccagcttacaaacagggacagaaccag ctgtataacgagctgaatctgggccgccga gaggaatatgacgtgctggataagcggagaggacgcgaccccgaaatgggaggcaagccc aggcgcaaaaaccctcaggaagg cctgtataacgagctgcagaaggacaaaatggcagaagcctattctgagatcggcatgaa gggggagcgacggagaggcaaagg gcacgatgggctgtaccagggactgagcaccgccacaaaggacacctatgatgctctgca tatgcaggcactgcctccaagg (SEQ ID NO: 14660). The 4-1BB costimulatory domain may comprise an amino acid sequence comprising KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL (SEQ ID NO: 14661) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

I^GRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL (SEQ ID NO: 14661) . The 4- IBB costimulatory domain may be encoded by the nucleic acid sequence comprising aagagaggcaggaagaaactgctgtatattttcaaacagcccttcatgcgccccgtgcag actacccaggaggaagacgggtgctcc tgtcgattccctgaggaagaggaaggcgggtgtgagctg (SEQ ID NO: 14662). The 4-1BB costimulatory domain may be located between the transmembrane domain and the CD28 costimulatory domain.

[0162] In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8a, IgG4, and/or CD4 sequence. In certain embodiments of the CARs of the disclosure, the hinge may comprise a sequence derived from a human CD8a sequence. The hinge may comprise a human CD8a amino acid sequence comprising TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 14663) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD (SEQ ID NO: 14663). The human CD8a hinge amino acid sequence may be encoded by the nucleic acid sequence comprising

actaccacaccagcacctagaccaccaactccagctccaaccatcgcgagtcagccc ctgagtctgagacctgaggcctgcaggcc agctgcaggaggagctgtgcacaccaggggcctggacttcgcctgcgac (SEQ ID NO: 14664).

ScFv

[0163] The disclosure provides single chain variable fragment (scFv) compositions and methods for use of these compositions to recognize and bind to a specific target protein. ScFv compositions comprise a heavy chain variable region and a light chain variable region of an antibody. ScFv compositions may be incorporated into an antigen recognition region of a chimeric antigen receptor of the disclosure. ScFvs are fusion proteins of the variable regions of the heavy (VH) and light (VL) chains of immunoglobulins, and the VH and VL domains are connected with a short peptide linker. ScFvs retain the specificity of the original immunoglobulin, despite removal of the constant regions and the introduction of the linker. An exemplary linker comprises a sequence of GGGGSGGGGSGGGGS (SEQ ID NO:

14665).

Centyrins

[0164] Centyrins of the disclosure specifically bind to an antigen. Chimeric antigen receptors of the disclosure comprising one or more Centyrins that specifically bind an antigen may be used to direct the specificity of a cell, (e.g. a cytotoxic immune cell) towards the specific antigen.

[0165] Centyrins of the disclosure may comprise a protein scaffold, wherein the scaffold is capable of specifically binding an antigen. Centyrins of the disclosure may comprise a protein scaffold comprising a consensus sequence of at least one fibronectin type III (FN3) domain, wherein the scaffold is capable of specifically binding an antigen. The at least one fibronectin type III (FN3) domain may be derived from a human protein. The human protein may be Tenascin-C. The consensus sequence may comprise

LPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTVPGSERSYDL TGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT (SEQ ID NO: 14488) or

MLPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTVPGSERSY D LTGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT (SEQ ID NO: 14489). The consensus sequence may comprise an amino sequence at least 74% identical to

LPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTVPGSERSYDL TGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT (SEQ ID NO: 14488) or

MLPAPKNLVVSEVTEDSLRLSWTAPDAAFDSFLIQYQESEKVGEAINLTVPGSERSY D LTGLKPGTEYTVSIYGVKGGHRSNPLSAEFTT (SEQ ID NO: 14489). The consensus sequence may encoded by a nucleic acid sequence comprising

atgctgcctgcaccaaagaacctggtggtgtctcatgtgacagaggatagtgccaga ctgtcatggactgctcccgacgcagccttcg atagttttatcatcgtgtaccgggagaacatcgaaaccggcgaggccattgtcctgacag tgccagggtccgaacgctcttatgacctg acagatctgaagcccggaactgagtactatgtgcagatcgccggcgtcaaaggaggcaat atcagcttccctctgtccgcaatcttcac caca (SEQ ID NO: 14490). The consensus sequence may be modified at one or more positions within (a) a A-B loop comprising or consisting of the amino acid residues TEDS (SEQ ID NO: 14491) at positions 13-16 of the consensus sequence; (b) a B-C loop comprising or consisting of the amino acid residues TAPDAAF (SEQ ID NO: 14492) at positions 22-28 of the consensus sequence; (c) a C-D loop comprising or consisting of the amino acid residues SEKVGE (SEQ ID NO: 14493) at positions 38-43 of the consensus sequence; (d) a D-E loop comprising or consisting of the amino acid residues GSER (SEQ ID NO: 14494) at positions 51-54 of the consensus sequence; (e) a E-F loop comprising or consisting of the amino acid residues GLKPG (SEQ ID NO: 14495) at positions 60-64 of the consensus sequence; (f) a F-G loop comprising or consisting of the amino acid residues KGGHRSN (SEQ ID NO: 14496) at positions 75-81 of the consensus sequence; or (g) any combination of (a)-(f). Centyrins of the disclosure may comprise a consensus sequence of at least 5 fibronectin type III (FN3) domains, at least 10 fibronectin type III (FN3) domains or at least 15 fibronectin type III (FN3) domains. The scaffold may bind an antigen with at least one affinity selected from a KD of less than or equal to 10 ^~9 M, less than or equal to 10 ^~10 M, less than or equal to 10 ^~n M, less than or equal to 10 ^~12 M, less than or equal to 10 ^~1 M, less than or equal to 10 ^~14 M, and less than or equal to 10 ^~15 M. The KD may be determined by surface plasmon resonance.

[0166] The term "antibody mimetic" is intended to describe an organic compound that specifically binds a target sequence and has a structure distinct from a naturally-occurring antibody. Antibody mimetics may comprise a protein, a nucleic acid, or a small molecule. The target sequence to which an antibody mimetic of the disclosure specifically binds may be an antigen. Antibody mimetics may provide superior properties over antibodies including, but not limited to, superior solubility, tissue penetration, stability towards heat and enzymes (e.g. resistance to enzymatic degradation), and lower production costs. Exemplary antibody mimetics include, but are not limited to, an affibody, an afflilin, an affimer, an affitin, an alphabody, an anticalin, and avimer (also known as avidity multimer), a DARPin (Designed Ankyrin Repeat Protein), a Fynomer, a Kunitz domain peptide, and a monobody.

[0167] Affibody molecules of the disclosure comprise a protein scaffold comprising or consisting of one or more alpha helix without any disulfide bridges. Preferably, affibody molecules of the disclosure comprise or consist of three alpha helices. For example, an affibody molecule of the disclosure may comprise an immunoglobulin binding domain. An affibody molecule of the disclosure may comprise the Z domain of protein A.

[0168] Affilin molecules of the disclosure comprise a protein scaffold produced by modification of exposed amino acids of, for example, either gamma-B crystallin or ubiquitin. Affilin molecules functionally mimic an antibody's affinity to antigen, but do not structurally mimic an antibody. In any protein scaffold used to make an affilin, those amino acids that are accessible to solvent or possible binding partners in a properly-folded protein molecule are considered exposed amino acids. Any one or more of these exposed amino acids may be modified to specifically bind to a target sequence or antigen.

[0169] Affimer molecules of the disclosure comprise a protein scaffold comprising a highly stable protein engineered to display peptide loops that provide a high affinity binding site for a specific target sequence. Exemplary affimer molecules of the disclosure comprise a protein scaffold based upon a cystatin protein or tertiary structure thereof. Exemplary affimer molecules of the disclosure may share a common tertiary structure of comprising an alpha- helix lying on top of an anti-parallel beta-sheet.

[0170] Affitin molecules of the disclosure comprise an artificial protein scaffold, the structure of which may be derived, for example, from a DNA binding protein (e.g. the DNA binding protein Sac7d). Affitins of the disclosure selectively bind a target sequence, which may be the entirety or part of an antigen. Exemplary affitins of the disclosure are

manufactured by randomizing one or more amino acid sequences on the binding surface of a DNA binding protein and subjecting the resultant protein to ribosome display and selection. Target sequences of affitins of the disclosure may be found, for example, in the genome or on the surface of a peptide, protein, virus, or bacteria. In certain embodiments of the disclosure, an affitin molecule may be used as a specific inhibitor of an enzyme. Affitin molecules of the disclosure may include heat-resistant proteins or derivatives thereof.

[0171] Alphabody molecules of the disclosure may also be referred to as Cell-Penetrating Alphabodies (CPAB). Alphabody molecules of the disclosure comprise small proteins (typically of less than 10 kDa) that bind to a variety of target sequences (including antigens). Alphabody molecules are capable of reaching and binding to intracellular target sequences. Structurally, alphabody molecules of the disclosure comprise an artificial sequence forming single chain alpha helix (similar to naturally occurring coiled-coil structures). Alphabody molecules of the disclosure may comprise a protein scaffold comprising one or more amino acids that are modified to specifically bind target proteins. Regardless of the binding specificity of the molecule, alphabody molecules of the disclosure maintain correct folding and thermostability.

[0172] Anticalin molecules of the disclosure comprise artificial proteins that bind to target sequences or sites in either proteins or small molecules. Anticalin molecules of the disclosure may comprise an artificial protein derived from a human lipocalin. Anticalin molecules of the disclosure may be used in place of, for example, monoclonal antibodies or fragments thereof. Anticalin molecules may demonstrate superior tissue penetration and thermostability than monoclonal antibodies or fragments thereof. Exemplary anticalin molecules of the disclosure may comprise about 180 amino acids, having a mass of approximately 20 kDa. Structurally, anticalin molecules of the disclosure comprise a barrel structure comprising antiparallel beta-strands pairwise connected by loops and an attached alpha helix. In preferred embodiments, anticalin molecules of the disclosure comprise a barrel structure comprising eight antiparallel beta-strands pairwise connected by loops and an attached alpha helix.

[0173] Avimer molecules of the disclosure comprise an artificial protein that specifically binds to a target sequence (which may also be an antigen). Avimers of the disclosure may recognize multiple binding sites within the same target or within distinct targets. When an avimer of the disclosure recognize more than one target, the avimer mimics function of a bi- specific antibody. The artificial protein avimer may comprise two or more peptide sequences of approximately 30-35 amino acids each. These peptides may be connected via one or more linker peptides. Amino acid sequences of one or more of the peptides of the avimer may be derived from an A domain of a membrane receptor. Avimers have a rigid structure that may optionally comprise disulfide bonds and/or calcium. Avimers of the disclosure may demonstrate greater heat stability compared to an antibody.

[0174] DARPins (Designed Ankyrin Repeat Proteins) of the disclosure comprise genetically-engineered, recombinant, or chimeric proteins having high specificity and high affinity for a target sequence. In certain embodiments, DARPins of the disclosure are derived from ankyrin proteins and, optionally, comprise at least three repeat motifs (also referred to as repetitive structural units) of the ankyrin protein. Ankyrin proteins mediate high-affinity protein-protein interactions. DARPins of the disclosure comprise a large target interaction surface.

[0175] Fynomers of the disclosure comprise small binding proteins (about 7 kDa) derived from the human Fyn SH3 domain and engineered to bind to target sequences and molecules with equal affinity and equal specificity as an antibody.

[0176] Kunitz domain peptides of the disclosure comprise a protein scaffold comprising a Kunitz domain. Kunitz domains comprise an active site for inhibiting protease activity. Structurally, Kunitz domains of the disclosure comprise a disulfide-rich alpha+beta fold. This structure is exemplified by the bovine pancreatic trypsin inhibitor. Kunitz domain peptides recognize specific protein structures and serve as competitive protease inhibitors. Kunitz domains of the disclosure may comprise Ecallantide (derived from a human lipoprotein- associated coagulation inhibitor (LACI)).

[0177] Monobodies of the disclosure are small proteins (comprising about 94 amino acids and having a mass of about 10 kDa) comparable in size to a single chain antibody. These genetically engineered proteins specifically bind target sequences including antigens.

Monobodies of the disclosure may specifically target one or more distinct proteins or target sequences. In preferred embodiments, monobodies of the disclosure comprise a protein scaffold mimicking the structure of human fibronectin, and more preferably, mimicking the structure of the tenth extracellular type III domain of fibronectin. The tenth extracellular type III domain of fibronectin, as well as a monobody mimetic thereof, contains seven beta sheets forming a barrel and three exposed loops on each side corresponding to the three complementarity determining regions (CDRs) of an antibody. In contrast to the structure of the variable domain of an antibody, a monobody lacks any binding site for metal ions as well as a central disulfide bond. Multispecific monobodies may be optimized by modifying the loops BC and FG. Monobodies of the disclosure may comprise an adnectin.

VHH

[0178] In certain embodiments, the CAR comprises a single domain antibody (SdAb). In certain embodiments, the SdAb is a VHH.

[0179] The disclosure provides chimeric antigen receptors (CARs) comprising at least one VHH (a VCAR). Chimeric antigen receptors of the disclosure may comprise more than one VHH. For example, a bi-specific VCAR may comprise two VHHs that specifically bind two distinct antigens.

[0180] VHH proteins of the disclosure specifically bind to an antigen. Chimeric antigen receptors of the disclosure comprising one or more VHHs that specifically bind an antigen may be used to direct the specificity of a cell, (e.g. a cytotoxic immune cell) towards the specific antigen.

[0181] At least one VHH protein or VCAR of the disclosure can be optionally produced by a cell line, a mixed cell line, an immortalized cell or clonal population of immortalized cells, as well known in the art. See, e.g., Ausubel, et al, ed., Current Protocols in Molecular Biology, John Wiley & Sons, Inc., NY, N.Y. (1987-2001); Sambrook, et al, Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor, N.Y. (1989); Harlow and Lane, Antibodies, a Laboratory Manual, Cold Spring Harbor, N.Y. (1989); Colligan, et al, eds., Current Protocols in Immunology, John Wiley & Sons, Inc., NY (1994-2001); Colligan et al, Current Protocols in Protein Science, John Wiley & Sons, NY, N.Y., (1997-2001).

[0182] Amino acids from a VHH protein can be altered, added and/or deleted to reduce immunogenicity or reduce, enhance or modify binding, affinity, on-rate, off-rate, avidity, specificity, half-life, stability, solubility or any other suitable characteristic, as known in the art.

[0183] Optionally, VHH proteins can be engineered with retention of high affinity for the antigen and other favorable biological properties. To achieve this goal, the VHH proteins can be optionally prepared by a process of analysis of the parental sequences and various conceptual engineered products using three-dimensional models of the parental and engineered sequences. Three-dimensional models are commonly available and are familiar to those skilled in the art. Computer programs are available which illustrate and display probable three-dimensional conformational structures of selected candidate sequences and can measure possible immunogenicity (e.g., Immunofilter program of Xencor, Inc. of Monrovia, Calif). Inspection of these displays permits analysis of the likely role of the residues in the functioning of the candidate sequence, i.e., the analysis of residues that influence the ability of the candidate VHH protein to bind its antigen. In this way, residues can be selected and combined from the parent and reference sequences so that the desired characteristic, such as affinity for the target antigen(s), is achieved. Alternatively, or in addition to, the above procedures, other suitable methods of engineering can be used.

[0184] Screening VHH for specific binding to similar proteins or fragments can be conveniently achieved using nucleotide (DNA or RNA display) or peptide display libraries, for example, in vitro display. This method involves the screening of large collections of peptides for individual members having the desired function or structure. The displayed nucleotide or peptide sequences can be from 3 to 5000 or more nucleotides or amino acids in length, frequently from 5-100 amino acids long, and often from about 8 to 25 amino acids long. In addition to direct chemical synthetic methods for generating peptide libraries, several recombinant DNA methods have been described. One type involves the display of a peptide sequence on the surface of a bacteriophage or cell. Each bacteriophage or cell contains the nucleotide sequence encoding the particular displayed peptide sequence. The VHH proteins of the disclosure can bind human or other mammalian proteins with a wide range of affinities (KD). In a preferred embodiment, at least one VHH of the present invention can optionally bind to a target protein with high affinity, for example, with a KD equal to or less than about 10 ^~7 M, such as but not limited to, 0.1-9.9 (or any range or value therein) X 10 ^~8 , 10 ^~9 , 10 ^~10 , 10 ^"11 , 10 ^~12 , 10 ^~13 , 10 ^~14 , 10 ^~15 or any range or value therein, as determined by surface plasmon resonance or the Kinexa method, as practiced by those of skill in the art.

[0185] The affinity or avidity of a VHH or a VCAR for an antigen can be determined experimentally using any suitable method. (See, for example, Berzofsky, et al, "Antibody- Antigen Interactions," In Fundamental Immunology, Paul, W. E., Ed., Raven Press: New York, N.Y. (1984); Kuby, Janis Immunology, W.H. Freeman and Company: New York, N.Y. (1992); and methods described herein). The measured affinity of a particular VHH-antigen or VCAR-antigen interaction can vary if measured under different conditions (e.g., salt concentration, pH). Thus, measurements of affinity and other antigen-binding parameters (e.g., KD, Kon, Kofi) are preferably made with standardized solutions of VHH or VCAR and antigen, and a standardized buffer, such as the buffer described herein.

[0186] Competitive assays can be performed with the VHH or VCAR of the disclosure in order to determine what proteins, antibodies, and other antagonists compete for binding to a target protein with the VHH or VCAR of the present invention and/or share the epitope region. These assays as readily known to those of ordinary skill in the art evaluate competition between antagonists or ligands for a limited number of binding sites on a protein. The protein and/or antibody is immobilized or insolubilized before or after the competition and the sample bound to the target protein is separated from the unbound sample, for example, by decanting (where the protein/antibody was preinsolubilized) or by centrifuging (where the protein/antibody was precipitated after the competitive reaction). Also, the competitive binding may be determined by whether function is altered by the binding or lack of binding of the VHH or VCAR to the target protein, e.g., whether the VCAR molecule inhibits or potentiates the enzymatic activity of, for example, a label. ELISA and other functional assays may be used, as well known in the art.

VH

[0187] In certain embodiments, the CAR comprises a single domain antibody (SdAb). In certain embodiments, the SdAb is a VH.

[0188] The disclosure provides chimeric antigen receptors (CARs) comprising a single domain antibody (VCARs). In certain embodiments, the single domain antibody comprises a VH. In certain embodiments, the VH is isolated or derived from a human sequence. In certain embodiments, VH comprises a human CDR sequence and/or a human framework sequence and a non-human or humanized sequence (e.g. a rat Fc domain). In certain embodiments, the VH is a fully humanized VH. In certain embodiments, the VH s neither a naturally occurring antibody nor a fragment of a naturally occurring antibody. In certain embodiments, the VH is not a fragment of a monoclonal antibody. In certain embodiments, the VH is a UniDab™ antibody (TeneoBio).

[0189] In certain embodiments, the VH is fully engineered using the UniRat™ (TeneoBio) system and "NGS-based Discovery" to produce the VH. Using this method, the specific VH are not naturally-occurring and are generated using fully engineered systems. The VH are not derived from naturally-occurring monoclonal antibodies (mAbs) that were either isolated directly from the host (for example, a mouse, rat or human) or directly from a single clone of cells or cell line (hybridoma). These VHs were not subsequently cloned from said cell lines. Instead, VH sequences are fully-engineered using the UniRat™ system as transgenes that comprise human variable regions (VH domains) with a rat Fc domain, and are thus human/rat chimeras without a light chain and are unlike the standard mAb format. The native rat genes are knocked out and the only antibodies expressed in the rat are from transgenes with VH domains linked to a Rat Fc (UniAbs). These are the exclusive Abs expressed in the UniRat. Next generation sequencing (NGS) and bioinformatics are used to identify the full antigen-specific repertoire of the heavy-chain antibodies generated by UniRat™ after immunization. Then, a unique gene assembly method is used to convert the antibody repertoire sequence information into large collections of fully -human heavy-chain antibodies that can be screened in vitro for a variety of functions. In certain embodiments, fully humanized VH are generated by fusing the human VH domains with human Fes in vitro (to generate a non-naturally occurring recombinant VH antibody). In certain embodiments, the VH are fully humanized, but they are expressed in vivo as human/rat chimera (human VH, rat Fc) without a light chain. Fully humanized VHs are expressed in vivo as human/rat chimera (human VH, rat Fc) without a light chain are about 80kDa (vs 150 kDa).

[0190] VCARs of the disclosure may comprise at least one VH of the disclosure. In certain embodiments, the VH of the disclosure may be modified to remove an Fc domain or a portion thereof. In certain embodiments, a framework sequence of the VH of the disclosure may be modified to, for example, improve expression, decrease immunogenicity or to improve function.

[0191] As used throughout the disclosure, the singular forms "a," "and," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a method" includes a plurality of such methods and reference to "a dose" includes reference to one or more doses and equivalents thereof known to those skilled in the art, and so forth.

[0192] The term "about" or "approximately" means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, "about" can mean within 1 or more standard deviations. Alternatively, "about" can mean a range of up to 20%, or up to 10%, or up to 5%, or up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5 -fold, and more preferably within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term "about" meaning within an acceptable error range for the particular value should be assumed. [0193] The disclosure provides isolated or substantially purified polynucleotide or protein compositions. An "isolated" or "purified" polynucleotide or protein, or biologically active portion thereof, is substantially or essentially free from components that normally accompany or interact with the polynucleotide or protein as found in its naturally occurring environment. Thus, an isolated or purified polynucleotide or protein is substantially free of other cellular material or culture medium when produced by recombinant techniques, or substantially free of chemical precursors or other chemicals when chemically synthesized. Optimally, an "isolated" polynucleotide is free of sequences (optimally protein encoding sequences) that naturally flank the polynucleotide (i.e., sequences located at the 5' and 3' ends of the polynucleotide) in the genomic DNA of the organism from which the polynucleotide is derived. For example, in various embodiments, the isolated polynucleotide can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb of nucleotide sequence that naturally flank the polynucleotide in genomic DNA of the cell from which the polynucleotide is derived. A protein that is substantially free of cellular material includes preparations of protein having less than about 30%, 20%, 10%, 5%, or 1% (by dry weight) of contaminating protein. When the protein of the invention or biologically active portion thereof is recombinantly produced, optimally culture medium represents less than about 30%, 20%, 10%, 5%, or 1 % (by dry weight) of chemical precursors or non-protein-of-interest chemicals.

[0194] The disclosure provides fragments and variants of the disclosed DNA sequences and proteins encoded by these DNA sequences. As used throughout the disclosure, the term "fragment" refers to a portion of the DNA sequence or a portion of the amino acid sequence and hence protein encoded thereby. Fragments of a DNA sequence comprising coding sequences may encode protein fragments that retain biological activity of the native protein and hence DNA recognition or binding activity to a target DNA sequence as herein described. Alternatively, fragments of a DNA sequence that are useful as hybridization probes generally do not encode proteins that retain biological activity or do not retain promoter activity. Thus, fragments of a DNA sequence may range from at least about 20 nucleotides, about 50 nucleotides, about 100 nucleotides, and up to the full-length polynucleotide of the invention.

[0195] Nucleic acids or proteins of the disclosure can be constructed by a modular approach including preassembling monomer units and/or repeat units in target vectors that can subsequently be assembled into a final destination vector. Polypeptides of the disclosure may comprise repeat monomers of the disclosure and can be constructed by a modular approach by preassembling repeat units in target vectors that can subsequently be assembled into a final destination vector. The disclosure provides polypeptide produced by this method as well nucleic acid sequences encoding these polypeptides. The disclosure provides host organisms and cells comprising nucleic acid sequences encoding polypeptides produced this modular approach.

[0196] The term "antibody" is used in the broadest sense and specifically covers single monoclonal antibodies (including agonist and antagonist antibodies) and antibody compositions with polyepitopic specificity. It is also within the scope hereof to use natural or synthetic analogs, mutants, variants, alleles, homologs and orthologs (herein collectively referred to as "analogs") of the antibodies hereof as defined herein. Thus, according to one embodiment hereof, the term "antibody hereof in its broadest sense also covers such analogs. Generally, in such analogs, one or more amino acid residues may have been replaced, deleted and/or added, compared to the antibodies hereof as defined herein.

[0197] "Antibody fragment", and all grammatical variants thereof, as used herein are defined as a portion of an intact antibody comprising the antigen binding site or variable region of the intact antibody, wherein the portion is free of the constant heavy chain domains (i.e. CH2, CH3, and CH4, depending on antibody isotype) of the Fc region of the intact antibody.

Examples of antibody fragments include Fab, Fab', Fab'- SH, F(ab')2, and Fv fragments; diabodies; any antibody fragment that is a polypeptide having a primary structure consisting of one uninterrupted sequence of contiguous amino acid residues (referred to herein as a "single-chain antibody fragment" or "single chain polypeptide"), including without limitation (1) single-chain Fv (scFv) molecules (2) single chain polypeptides containing only one light chain variable domain, or a fragment thereof that contains the three CDRs of the light chain variable domain, without an associated heavy chain moiety and (3) single chain polypeptides containing only one heavy chain variable region, or a fragment thereof containing the three CDRs of the heavy chain variable region, without an associated light chain moiety; and multispecific or multivalent structures formed from antibody fragments. In an antibody fragment comprising one or more heavy chains, the heavy chain(s) can contain any constant domain sequence (e.g. CHI in the IgG isotype) found in a non-Fc region of an intact antibody, and/or can contain any hinge region sequence found in an intact antibody, and/or can contain a leucine zipper sequence fused to or situated in the hinge region sequence or the constant domain sequence of the heavy chain(s). The term further includes single domain antibodies ("sdAB") which generally refers to an antibody fragment having a single monomeric variable antibody domain, (for example, from camelids). Such antibody fragment types will be readily understood by a person having ordinary skill in the art. [0198] "Binding" refers to a sequence-specific, non-covalent interaction between

macromolecules (e.g., between a protein and a nucleic acid). Not all components of a binding interaction need be sequence-specific (e.g., contacts with phosphate residues in a DNA backbone), as long as the interaction as a whole is sequence-specific.

[0199] The term "comprising" is intended to mean that the compositions and methods include the recited elements, but do not exclude others. "Consisting essentially of when used to define compositions and methods, shall mean excluding other elements of any essential significance to the combination when used for the intended purpose. Thus, a composition consisting essentially of the elements as defined herein would not exclude trace contaminants or inert carriers. "Consisting of shall mean excluding more than trace elements of other ingredients and substantial method steps. Embodiments defined by each of these transition terms are within the scope of this invention.

[0200] The term "epitope" refers to an antigenic determinant of a polypeptide. An epitope could comprise three amino acids in a spatial conformation, which is unique to the epitope. Generally, an epitope consists of at least 4, 5, 6, or 7 such amino acids, and more usually, consists of at least 8, 9, or 10 such amino acids. Methods of determining the spatial conformation of amino acids are known in the art, and include, for example, x-ray crystallography and two-dimensional nuclear magnetic resonance.

[0201] As used herein, "expression" refers to the process by which polynucleotides are transcribed into mRNA and/or the process by which the transcribed mRNA is subsequently being translated into peptides, polypeptides, or proteins. If the polynucleotide is derived from genomic DNA, expression may include splicing of the mRNA in a eukaryotic cell.

[0202] "Gene expression" refers to the conversion of the information, contained in a gene, into a gene product. A gene product can be the direct transcriptional product of a gene (e.g., mRNA, tRNA, rRNA, antisense RNA, ribozyme, shRNA, micro RNA, structural RNA or any other type of RNA) or a protein produced by translation of an mRNA. Gene products also include RNAs which are modified, by processes such as capping, polyadenylation, methylation, and editing, and proteins modified by, for example, methylation, acetylation, phosphorylation, ubiquitination, ADP-ribosylation, myristilation, and glycosylation.

[0203] "Modulation" or "regulation" of gene expression refers to a change in the activity of a gene. Modulation of expression can include, but is not limited to, gene activation and gene repression. [0204] The term "operatively linked" or its equivalents (e.g., "linked operatively") means two or more molecules are positioned with respect to each other such that they are capable of interacting to affect a function attributable to one or both molecules or a combination thereof.

[0205] Non-covalently linked components and methods of making and using non-covalently linked components, are disclosed. The various components may take a variety of different forms as described herein. For example, non-covalently linked (i.e., operatively linked) proteins may be used to allow temporary interactions that avoid one or more problems in the art. The ability of non-covalently linked components, such as proteins, to associate and dissociate enables a functional association only or primarily under circumstances where such association is needed for the desired activity. The linkage may be of duration sufficient to allow the desired effect.

[0206] A method for directing proteins to a specific locus in a genome of an organism is disclosed. The method may comprise the steps of providing a DNA localization component and providing an effector molecule, wherein the DNA localization component and the effector molecule are capable of operatively linking via a non-covalent linkage.

[0207] The term "scFv" refers to a single-chain variable fragment. scFv is a fusion protein of the variable regions of the heavy (VH) and light chains (VL) of immunoglobulins, connected with a linker peptide. The linker peptide may be from about 5 to 40 amino acids or from about 10 to 30 amino acids or about 5, 10, 15, 20, 25, 30, 35, or 40 amino acids in length. Single-chain variable fragments lack the constant Fc region found in complete antibody molecules, and, thus, the common binding sites (e.g., Protein G) used to purify antibodies. The term further includes a scFv that is an intrabody, an antibody that is stable in the cytoplasm of the cell, and which may bind to an intracellular protein.

[0208] The term "single domain antibody" means an antibody fragment having a single monomeric variable antibody domain which is able to bind selectively to a specific antigen. A single-domain antibody generally is a peptide chain of about 110 amino acids long, comprising one variable domain (VH) of a heavy-chain antibody, or of a common IgG, which generally have similar affinity to antigens as whole antibodies, but are more heat-resistant and stable towards detergents and high concentrations of urea. Examples are those derived from camelid or fish antibodies. Alternatively, single-domain antibodies can be made from common murine or human IgG with four chains.

[0209] The terms "specifically bind" and "specific binding" as used herein refer to the ability of an antibody, an antibody fragment or a nanobody to preferentially bind to a particular antigen that is present in a homogeneous mixture of different antigens. In certain embodiments, a specific binding interaction will discriminate between desirable and undesirable antigens in a sample. In certain embodiments more than about ten- to 100-fold or more (e.g., more than about 1000- or 10,000-fold). "Specificity" refers to the ability of an immunoglobulin or an immunoglobulin fragment, such as a nanobody, to bind preferentially to one antigenic target versus a different antigenic target and does not necessarily imply high affinity.

[0210] A "target site" or "target sequence" is a nucleic acid sequence that defines a portion of a nucleic acid to which a binding molecule will bind, provided sufficient conditions for binding exist.

[0211] The terms "nucleic acid" or "oligonucleotide" or "polynucleotide" refer to at least two nucleotides covalently linked together. The depiction of a single strand also defines the sequence of the complementary strand. Thus, a nucleic acid may also encompass the complementary strand of a depicted single strand. A nucleic acid of the disclosure also encompasses substantially identical nucleic acids and complements thereof that retain the same structure or encode for the same protein.

[0212] Probes of the disclosure may comprise a single stranded nucleic acid that can hybridize to a target sequence under stringent hybridization conditions. Thus, nucleic acids of the disclosure may refer to a probe that hybridizes under stringent hybridization conditions.

[0213] Nucleic acids of the disclosure may be single- or double-stranded. Nucleic acids of the disclosure may contain double-stranded sequences even when the majority of the molecule is single-stranded. Nucleic acids of the disclosure may contain single-stranded sequences even when the majority of the molecule is double-stranded. Nucleic acids of the disclosure may include genomic DNA, cDNA, RNA, or a hybrid thereof. Nucleic acids of the disclosure may contain combinations of deoxyribo- and ribo-nucleotides. Nucleic acids of the disclosure may contain combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine and isoguanine. Nucleic acids of the disclosure may be synthesized to comprise non-natural amino acid modifications. Nucleic acids of the disclosure may be obtained by chemical synthesis methods or by recombinant methods.

[0214] Nucleic acids of the disclosure, either their entire sequence, or any portion thereof, may be non-naturally occurring. Nucleic acids of the disclosure may contain one or more mutations, substitutions, deletions, or insertions that do not naturally-occur, rendering the entire nucleic acid sequence non-naturally occurring. Nucleic acids of the disclosure may contain one or more duplicated, inverted or repeated sequences, the resultant sequence of which does not naturally-occur, rendering the entire nucleic acid sequence non-naturally occurring. Nucleic acids of the disclosure may contain modified, artificial, or synthetic nucleotides that do not naturally-occur, rendering the entire nucleic acid sequence non- naturally occurring.

[0215] Given the redundancy in the genetic code, a plurality of nucleotide sequences may encode any particular protein. All such nucleotides sequences are contemplated herein.

[0216] As used throughout the disclosure, the term "operably linked" refers to the expression of a gene that is under the control of a promoter with which it is spatially connected. A promoter can be positioned 5' (upstream) or 3' (downstream) of a gene under its control. The distance between a promoter and a gene can be approximately the same as the distance between that promoter and the gene it controls in the gene from which the promoter is derived. Variation in the distance between a promoter and a gene can be accommodated without loss of promoter function.

[0217] As used throughout the disclosure, the term "promoter" refers to a synthetic or naturally-derived molecule which is capable of conferring, activating or enhancing expression of a nucleic acid in a cell. A promoter can comprise one or more specific transcriptional regulatory sequences to further enhance expression and/or to alter the spatial expression and/or temporal expression of same. A promoter can also comprise distal enhancer or repressor elements, which can be located as much as several thousand base pairs from the start site of transcription. A promoter can be derived from sources including viral, bacterial, fungal, plants, insects, and animals. A promoter can regulate the expression of a gene component constitutively or differentially with respect to cell, the tissue or organ in which expression occurs or, with respect to the developmental stage at which expression occurs, or in response to external stimuli such as physiological stresses, pathogens, metal ions, or inducing agents. Representative examples of promoters include the bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator-promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV IE promoter, EF-1 Alpha promoter, CAG promoter, SV40 early promoter or SV40 late promoter and the CMV IE promoter.

[0218] As used throughout the disclosure, the term "substantially complementary" refers to a first sequence that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical to the complement of a second sequence over a region of 8, 9, 10, 11, 12, 13, 14, 15,

16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540, or more nucleotides or amino acids, or that the two sequences hybridize under stringent hybridization conditions.

[0219] As used throughout the disclosure, the term "substantially identical" refers to a first and second sequence are at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% or 99% identical over a region of 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, 450, 540 or more nucleotides or amino acids, or with respect to nucleic acids, if the first sequence is substantially complementary to the complement of the second sequence.

[0220] As used throughout the disclosure, the term "variant" when used to describe a nucleic acid, refers to (i) a portion or fragment of a referenced nucleotide sequence; (ii) the complement of a referenced nucleotide sequence or portion thereof; (iii) a nucleic acid that is substantially identical to a referenced nucleic acid or the complement thereof; or (iv) a nucleic acid that hybridizes under stringent conditions to the referenced nucleic acid, complement thereof, or a sequences substantially identical thereto.

[0221] As used throughout the disclosure, the term "vector" refers to a nucleic acid sequence containing an origin of replication. A vector can be a viral vector, bacteriophage, bacterial artificial chromosome or yeast artificial chromosome. A vector can be a DNA or RNA vector. A vector can be a self-replicating extrachromosomal vector, and preferably, is a DNA plasmid. A vector may comprise a combination of an amino acid with a DNA sequence, an RNA sequence, or both a DNA and an RNA sequence.

[0222] As used throughout the disclosure, the term "variant" when used to describe a peptide or polypeptide, refers to a peptide or polypeptide that differs in amino acid sequence by the insertion, deletion, or conservative substitution of amino acids, but retain at least one biological activity. Variant can also mean a protein with an amino acid sequence that is substantially identical to a referenced protein with an amino acid sequence that retains at least one biological activity.

[0223] A conservative substitution of an amino acid, i.e., replacing an amino acid with a different amino acid of similar properties (e.g., hydrophilicity, degree and distribution of charged regions) is recognized in the art as typically involving a minor change. These minor changes can be identified, in part, by considering the hydropathic index of amino acids, as understood in the art. Kyte et al, J. Mol. Biol. 157: 105-132 (1982). The hydropathic index of an amino acid is based on a consideration of its hydrophobicity and charge. Amino acids of similar hydropathic indexes can be substituted and still retain protein function. In one aspect, amino acids having hydropathic indexes of ±2 are substituted. The hydrophilicity of amino acids can also be used to reveal substitutions that would result in proteins retaining biological function. A consideration of the hydrophilicity of amino acids in the context of a peptide permits calculation of the greatest local average hydrophilicity of that peptide, a useful measure that has been reported to correlate well with antigenicity and immunogenicity. U.S. Patent No. 4,554,101, incorporated fully herein by reference.

[0224] Substitution of amino acids having similar hydrophilicity values can result in peptides retaining biological activity, for example immunogenicity. Substitutions can be performed with amino acids having hydrophilicity values within ±2 of each other. Both the

hyrophobicity index and the hydrophilicity value of amino acids are influenced by the particular side chain of that amino acid. Consistent with that observation, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, and particularly the side chains of those amino acids, as revealed by the hydrophobicity, hydrophilicity, charge, size, and other properties.

[0225] As used herein, "conservative" amino acid substitutions may be defined as set out in Tables A, B, or C below. In some embodiments, fusion polypeptides and/or nucleic acids encoding such fusion polypeptides include conservative substitutions have been introduced by modification of polynucleotides encoding polypeptides of the invention. Amino acids can be classified according to physical properties and contribution to secondary and tertiary protein structure. A conservative substitution is a substitution of one amino acid for another amino acid that has similar properties. Exemplary conservative substitutions are set out in

Table A.

[0226] Table A ~ Conservative Substitutions I

[0227] Alternately, conservative amino acids can be grouped as described in Lehninger, (Biochemistry, Second Edition; Worth Publishers, Inc. NY, N.Y. (1975), pp. 71-77) as set forth in Table B. [0228] Table B ~ Conservative Substitutions II

[0229] Alternately, exemplary conservative substitutions are set out in Table C.

[0230] Table C ~ Conservative Substitutions III

Thr (T) Ser

Trp (W) Tyr Phe He

Tyr (Y) Trp Phe Thr Ser

Val (V) He Leu Met Ala

[0231] It should be understood that the polypeptides of the disclosure are intended to include polypeptides bearing one or more insertions, deletions, or substitutions, or any combination thereof, of amino acid residues as well as modifications other than insertions, deletions, or substitutions of amino acid residues. Polypeptides or nucleic acids of the disclosure may contain one or more conservative substitution.

[0232] As used throughout the disclosure, the term "more than one" of the aforementioned amino acid substitutions refers to 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1, 12, 13, 14, 15, 16, 17, 18, 19, or 20 or more of the recited amino acid substitutions. The term "more than one" may refer to 2, 3, 4, or 5 of the recited amino acid substitutions.

[0233] Polypeptides and proteins of the disclosure, either their entire sequence, or any portion thereof, may be non-naturally occurring. Polypeptides and proteins of the disclosure may contain one or more mutations, substitutions, deletions, or insertions that do not naturally-occur, rendering the entire amino acid sequence non-naturally occurring.

Polypeptides and proteins of the disclosure may contain one or more duplicated, inverted or repeated sequences, the resultant sequence of which does not naturally-occur, rendering the entire amino acid sequence non-naturally occurring. Polypeptides and proteins of the disclosure may contain modified, artificial, or synthetic amino acids that do not naturally- occur, rendering the entire amino acid sequence non-naturally occurring.

[0234] As used throughout the disclosure, "sequence identity" may be determined by using the stand-alone executable BLAST engine program for blasting two sequences (bl2seq), which can be retrieved from the National Center for Biotechnology Information (NCBI) ftp site, using the default parameters (Tatusova and Madden, FEMS Microbiol Lett., 1999, 174, 247-250; which is incorporated herein by reference in its entirety). The terms "identical" or "identity" when used in the context of two or more nucleic acids or polypeptide sequences, refer to a specified percentage of residues that are the same over a specified region of each of the sequences. The percentage can be calculated by optimally aligning the two sequences, comparing the two sequences over the specified region, determining the number of positions at which the identical residue occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the specified region, and multiplying the result by 100 to yield the percentage of sequence identity. In cases where the two sequences are of different lengths or the alignment produces one or more staggered ends and the specified region of comparison includes only a single sequence, the residues of single sequence are included in the denominator but not the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) can be considered equivalent. Identity can be performed manually or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

[0235] As used throughout the disclosure, the term "endogenous" refers to nucleic acid or protein sequence naturally associated with a target gene or a host cell into which it is introduced.

[0236] As used throughout the disclosure, the term "exogenous" refers to nucleic acid or protein sequence not naturally associated with a target gene or a host cell into which it is introduced, including non-naturally occurring multiple copies of a naturally occurring nucleic acid, e.g., DNA sequence, or naturally occurring nucleic acid sequence located in a non- naturally occurring genome location.

[0237] The disclosure provides methods of introducing a polynucleotide construct comprising a DNA sequence into a host cell. By "introducing" is intended presenting to the plant the polynucleotide construct in such a manner that the construct gains access to the interior of the host cell. The methods of the invention do not depend on a particular method for introducing a polynucleotide construct into a host cell, only that the polynucleotide construct gains access to the interior of one cell of the host. Methods for introducing polynucleotide constructs into bacteria, plants, fungi and animals are known in the art including, but not limited to, stable transformation methods, transient transformation methods, and virus-mediated methods.

Transposons/T ansposases

[0238] Exemplary transposon/transposase systems of the disclosure include, but are not limited to, piggyBac transposons and transposases, Sleeping Beauty transposons and transposases, Helraiser transposons and transposases and Tol2 transposons and transposases.

[0239] The piggyBac transposase recognizes transposon-specific inverted terminal repeat sequences (ITRs) on the ends of the transposon, and moves the contents between the ITRs into TTAA chromosomal sites. The piggyBac transposon system has no payload limit for the genes of interest that can be included between the ITRs. In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™ or a Super piggyBac™ (SPB) transposase. In certain

embodiments, and, in particular, those embodiments wherein the transposase is a Super piggyBac™ (SPB) transposase, the sequence encoding the transposase is an mRNA sequence.

[0240] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme. The piggyBac (PB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV

541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID

NO: 14487) .

[0241] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at one or more of positions 30, 165, 282, or 538 of the sequence:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV

541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID

NO: 14487) .

[0242] In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at two or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at three or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac™ (PB) transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at each of the following positions 30, 165, 282, and 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 165 of the sequence of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G). In certain embodiments, the amino acid substitution at position 282 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 538 of the sequence of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N).

[0243] In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) transposase enzyme. In certain embodiments, the Super piggyBac™ (SPB) transposase enzymes of the disclosure may comprise or consist of the amino acid sequence of the sequence of SEQ ID NO: 14487 wherein the amino acid substitution at position 30 is a substitution of a valine (V) for an isoleucine (I), the amino acid substitution at position 165 is a substitution of a serine (S) for a glycine (G), the amino acid substitution at position 282 is a substitution of a valine (V) for a methionine (M), and the amino acid substitution at position 538 is a substitution of a lysine (K) for an asparagine (N). In certain embodiments, the Super piggyBac™ (SPB) transposase enzyme may comprise or consist of an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ

361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC

421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO: 14484) .

[0244] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 3, 46, 82, 103, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 258, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 486, 503, 552, 570 and 591 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 46, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 485, 503, 552 and 570. In certain embodiments, the amino acid substitution at position 3 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for a serine (S). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an alanine (A). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 82 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 119 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for an arginine (R). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) a cysteine (C). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a

phenylalanine (F). In certain embodiments, the amino acid substitution at position 185 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 187 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for an alanine (A). In certain embodiments, the amino acid substitution at position 200 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a phenylalanine (F).In certain embodiments, the amino acid substitution at position 207 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a valine (V). In certain embodiments, the amino acid substitution at position 209 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a valine (V). In certain embodiments, the amino acid substitution at position 226 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a methionine (M). In certain embodiments, the amino acid substitution at position 235 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a leucine (L). In certain embodiments, the amino acid substitution at position 240 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 241 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 243 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a proline (P). In certain embodiments, the amino acid substitution at position 258 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a proline (P). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine for a proline (P). In certain embodiments, the amino acid substitution at position 315 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for an arginine (R).In certain embodiments, the amino acid substitution at position 319 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a threonine (T). In certain embodiments, the amino acid substitution at position 327 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 328 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a cysteine (C). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 421 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for the aspartic acid (D). In certain embodiments, the amino acid substitution at position 436 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a valine (V). In certain embodiments, the amino acid substitution at position 456 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a methionine (M). In certain embodiments, the amino acid substitution at position 470 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 485 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a serine (S). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a methionine (M). In certain embodiments, the amino acid substitution at position 552 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a glutamine (Q). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a glutamine (Q).

[0245] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at two, three, four, five, six or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 194 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 372 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for an arginine (R). In certain embodiments, the amino acid substitution at position 375 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a lysine (K). In certain embodiments, the amino acid substitution at position 450 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for an aspartic acid (D). In certain embodiments, the amino acid substitution at position 509 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a serine (S). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487. In certain embodiments, including those embodiments wherein the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, the piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 372, 375 and 450 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, and a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487. In certain embodiments, the piggyBac™ transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487 and a substitution of an asparagine (N) for an aspartic acid (D) at position 450 of SEQ ID NO: 14487.

[0246] The sleeping beauty transposon is transposed into the target genome by the Sleeping Beauty transposase that recognizes ITRs, and moves the contents between the ITRs into TA chromosomal sites. In various embodiments, SB transposon-mediated gene transfer, or gene transfer using any of a number of similar transposons, may be used in the compositions and methods of the disclosure.

[0247] In certain embodiments, and, in particular, those embodiments wherein the transposon is a Sleeping Beauty transposon, the transposase is a Sleeping Beauty transposase or a hyperactive Sleeping Beauty transposase (SBIOOX).

[0248] In certain embodiments of the methods of the disclosure, the Sleeping Beauty transposase enzyme comprises an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGKSKEISQD LRKKIVDLHK SGSSLGAISK RLKVPRSSVQ TIVRKYKHHG TTQPSYRSGR 61 RRVLSPRDER TLVRKVQINP RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGRSARKK 121 PLLQNRHKKA RLRFATAHGD KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN 181 TIPTVKHGGG SIMLWGCFAA GGTGALHKID GIMRKENYVD ILKQHLKTSV RKLKLGRKWV 241 FQMDNDPKHT SKWAKWLKD NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL 301 HQLCQEEWAK IHPTYCGKLV EGYPKRLTQV KQFKGNATKY (SEQ ID NO: 14485) .

[0249] In certain embodiments of the methods of the disclosure, the hyperactive Sleeping Beauty (SBIOOX) transposase enzyme comprises an amino acid sequence at least 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGKSKEISQD LRKRIVDLHK SGSSLGAISK RLAVPRSSVQ TIVRKYKHHG TTQPSYRSGR 61 RRVLSPRDER TLVRKVQINP RTTAKDLVKM LEETGTKVSI STVKRVLYRH NLKGHSARKK 121 PLLQNRHKKA RLRFATAHGD KDRTFWRNVL WSDETKIELF GHNDHRYVWR KKGEACKPKN 181 TIPTVKHGGG SIMLWGCFAA GGTGALHKID GIMDAVQYVD ILKQHLKTSV RKLKLGRKWV 241 FQHDNDPKHT SKWAKWLKD NKVKVLEWPS QSPDLNPIEN LWAELKKRVR ARRPTNLTQL 301 HQLCQEEWAK IHPNYCGKLV EGYPKRLTQV KQFKGNATKY (SEQ ID NO: 14486) .

[0250] The Helraiser transposon is transposed by the Helitron transposase. Helitron transposases mobilize the Helraiser transposon, an ancient element from the bat genome that was active about 30 to 36 million years ago. An exemplary Helraiser transposon of the disclosure includes Helibatl, which comprises a nucleic acid sequence comprising:

1 TCCTATATAA TAAAAGAGAA ACATGCAAAT TGACCATCCC TCCGCTACGC TCAAGCCACG

61 CCCACCAGCC AATCAGAAGT GACTATGCAA ATTAACCCAA CAAAGATGGC AGTTAAATTT

121 GCATACGCAG GTGTCAAGCG CCCCAGGAGG CAACGGCGGC CGCGGGCTCC CAGGACCTTC

181 GCTGGCCCCG GGAGGCGAGG CCGGCCGCGC CTAGCCACAC CCGCGGGCTC CCGGGACCTT

241 CGCCAGCAGA GAGCAGAGCG GGAGAGCGGG CGGAGAGCGG GAGGTTTGGA GGACTTGGCA

301 GAGCAGGAGG CCGCTGGACA TAGAGCAGAG CGAGAGAGAG GGTGGCTTGG AGGGCGTGGC

361 TCCCTCTGTC ACCCCAGCTT CCTCATCACA GCTGTGGAAA CTGACAGCAG GGAGGAGGAA

421 GTCCCACCCC CACAGAATCA GCCAGAATCA GCCGTTGGTC AGACAGCTCT CAGCGGCCTG

481 ACAGCCAGGA CTCTCATTCA CCTGCATCTC AGACCGTGAC AGTAGAGAGG TGGGACTATG

541 TCTAAAGAAC AACTGTTGAT ACAACGTAGC TCTGCAGCCG AAAGATGCCG GCGTTATCGA

601 CAGAAAATGT CTGCAGAGCA ACGTGCGTCT GATCTTGAAA GAAGGCGGCG CCTGCAACAG

661 AATGTATCTG AAGAGCAGCT ACTGGAAAAA CGTCGCTCTG AAGCCGAAAA ACAGCGGCGT

721 CATCGACAGA AAATGTCTAA AGACCAACGT GCCTTTGAAG TTGAAAGAAG GCGGTGGCGA

781 CGACAGAATA TGTCTAGAGA ACAGTCATCA ACAAGTACTA CCAATACCGG TAGGAACTGC

841 CTTCTCAGCA AAAATGGAGT ACATGAGGAT GCAATTCTCG AACATAGTTG TGGTGGAATG

901 ACTGTTCGAT GTGAATTTTG CCTATCACTA AATTTCTCTG ATGAAAAACC ATCCGATGGG

961 AAATTTACTC GATGTTGTAG CAAAGGGAAA GTCTGTCCAA ATGATATACA TTTTCCAGAT

1021 TACCCGGCAT ATTTAAAAAG ATTAATGACA AACGAAGATT CTGACAGTAA AAATTTCATG

1081 GAAAATATTC GTTCCATAAA TAGTTCTTTT GCTTTTGCTT CCATGGGTGC AAATATTGCA

1141 TCGCCATCAG GATATGGGCC ATACTGTTTT AGAATACACG GACAAGTTTA TCACCGTACT

1201 GGAACTTTAC ATCCTTCGGA TGGTGTTTCT CGGAAGTTTG CTCAACTCTA TATTTTGGAT

1261 ACAGCCGAAG CTACAAGTAA AAGATTAGCA ATGCCAGAAA ACCAGGGCTG CTCAGAAAGA

1321 CTCATGATCA ACATCAACAA CCTCATGCAT GAAATAAATG AATTAACAAA ATCGTACAAG

1381 ATGCTACATG AGGTAGAAAA GGAAGCCCAA TCTGAAGCAG CAGCAAAAGG TATTGCTCCC

1441 ACAGAAGTAA CAATGGCGAT TAAATACGAT CGTAACAGTG ACCCAGGTAG ATATAATTCT

1501 CCCCGTGTAA CCGAGGTTGC TGTCATATTC AGAAACGAAG ATGGAGAACC TCCTTTTGAA

1561 AGGGACTTGC TCATTCATTG TAAACCAGAT CCCAATAATC CAAATGCCAC TAAAATGAAA

1621 CAAATCAGTA TCCTGTTTCC TACATTAGAT GCAATGACAT ATCCTATTCT TTTTCCACAT

1681 GGTGAAAAAG GCTGGGGAAC AGATATTGCA TTAAGACTCA GAGACAACAG TGTAATCGAC

1741 AATAATACTA GACAAAATGT AAGGACACGA GTCACACAAA TGCAGTATTA TGGATTTCAT

1801 CTCTCTGTGC GGGACACGTT CAATCCTATT TTAAATGCAG GAAAATTAAC TCAACAGTTT

1861 ATTGTGGATT CATATTCAAA AATGGAGGCC AATCGGATAA ATTTCATCAA AGCAAACCAA

1921 TCTAAGTTGA GAGTTGAAAA ATATAGTGGT TTGATGGATT ATCTCAAATC TAGATCTGAA 1981 AATGACAATG TGCCGATTGG TAAAATGATA ATACTTCCAT CATCTTTTGA GGGTAGTCCC

2041 AGAAATATGC AGCAGCGATA TCAGGATGCT ATGGCAATTG TAACGAAGTA TGGCAAGCCC

2101 GATTTATTCA TAACCATGAC ATGCAACCCC AAATGGGCAG ATATTACAAA CAATTTACAA

2161 CGCTGGCAAA AAGTTGAAAA CAGACCTGAC TTGGTAGCCA GAGTTTTTAA TATTAAGCTG

2221 AATGCTCTTT TAAATGATAT ATGTAAATTC CATTTATTTG GCAAAGTAAT AGCTAAAATT

2281 CATGTCATTG AATTTCAGAA ACGCGGACTG CCTCACGCTC ACATATTATT GATATTAGAT

2341 AGTGAGTCCA AATTACGTTC AGAAGATGAC ATTGACCGTA TAGTTAAGGC AGAAATTCCA

2401 GATGAAGACC AGTGTCCTCG ACTTTTTCAA ATTGTAAAAT CAAATATGGT ACATGGACCA

2461 TGTGGAATAC AAAATCCAAA TAGTCCATGT ATGGAAAATG GAAAATGTTC AAAGGGATAT

2521 CCAAAAGAAT TTCAAAATGC GACCATTGGA AATATTGATG GATATCCCAA ATACAAACGA

2581 AGATCTGGTA GCACCATGTC TATTGGAAAT AAAGTTGTCG ATAACACTTG GATTGTCCCT

2641 TATAACCCGT ATTTGTGCCT TAAATATAAC TGTCATATAA ATGTTGAAGT CTGTGCATCA

2701 ATTAAAAGTG TCAAATATTT ATTTAAATAC ATCTATAAAG GGCACGATTG TGCAAATATT

2761 CAAATTTCTG AAAAAAATAT TATCAATCAT GACGAAGTAC AGGACTTCAT TGACTCCAGG

2821 TATGTGAGCG CTCCTGAGGC TGTTTGGAGA CTTTTTGCAA TGCGAATGCA TGACCAATCT

2881 CATGCAATCA CAAGATTAGC TATTCATTTG CCAAATGATC AGAATTTGTA TTTTCATACC

2941 GATGATTTTG CTGAAGTTTT AGATAGGGCT AAAAGGCATA ACTCGACTTT GATGGCTTGG

3001 TTCTTATTGA ATAGAGAAGA TTCTGATGCA CGTAATTATT ATTATTGGGA GATTCCACAG

3061 CATTATGTGT TTAATAATTC TTTGTGGACA AAACGCCGAA AGGGTGGGAA TAAAGTATTA

3121 GGTAGACTGT TCACTGTGAG CTTTAGAGAA CCAGAACGAT ATTACCTTAG ACTTTTGCTT

3181 CTGCATGTAA AAGGTGCGAT AAGTTTTGAG GATCTGCGAA CTGTAGGAGG TGTAACTTAT

3241 GATACATTTC ATGAAGCTGC TAAACACCGA GGATTATTAC TTGATGACAC TATCTGGAAA

3301 GATACGATTG ACGATGCAAT CATCCTTAAT ATGCCCAAAC AACTACGGCA ACTTTTTGCA

3361 TATATATGTG TGTTTGGATG TCCTTCTGCT GCAGACAAAT TATGGGATGA GAATAAATCT

3421 CATTTTATTG AAGATTTCTG TTGGAAATTA CACCGAAGAG AAGGTGCCTG TGTGAACTGT

3481 GAAATGCATG CCCTTAACGA AATTCAGGAG GTATTCACAT TGCATGGAAT GAAATGTTCA

3541 CATTTCAAAC TTCCGGACTA TCCTTTATTA ATGAATGCAA ATACATGTGA TCAATTGTAC

3601 GAGCAACAAC AGGCAGAGGT TTTGATAAAT TCTCTGAATG ATGAACAGTT GGCAGCCTTT

3661 CAGACTATAA CTTCAGCCAT CGAAGATCAA ACTGTACACC CCAAATGCTT TTTCTTGGAT

3721 GGTCCAGGTG GTAGTGGAAA AACATATCTG TATAAAGTTT TAACACATTA TATTAGAGGT

3781 CGTGGTGGTA CTGTTTTACC CACAGCATCT ACAGGAATTG CTGCAAATTT ACTTCTTGGT

3841 GGAAGAACCT TTCATTCCCA ATATAAATTA CCAATTCCAT TAAATGAAAC TTCAATTTCT

3901 AGACTCGATA TAAAGAGTGA AGTTGCTAAA ACCATTAAAA AGGCCCAACT TCTCATTATT

3961 GATGAATGCA CCATGGCATC CAGTCATGCT ATAAACGCCA TAGATAGATT ACTAAGAGAA

4021 ATTATGAATT TGAATGTTGC ATTTGGTGGG AAAGTTCTCC TTCTCGGAGG GGATTTTCGA

4081 CAATGTCTCA GTATTGTACC ACATGCTATG CGATCGGCCA TAGTACAAAC GAGTTTAAAG

4141 TACTGTAATG TTTGGGGATG TTTCAGAAAG TTGTCTCTTA AAACAAATAT GAGATCAGAG

4201 GATTCTGCTT ATAGTGAATG GTTAGTAAAA CTTGGAGATG GCAAACTTGA TAGCAGTTTT

4261 CATTTAGGAA TGGATATTAT TGAAATCCCC CATGAAATGA TTTGTAACGG ATCTATTATT

4321 GAAGCTACCT TTGGAAATAG TATATCTATA GATAATATTA AAAATATATC TAAACGTGCA

4381 ATTCTTTGTC CAAAAAATGA GCATGTTCAA AAATTAAATG AAGAAATTTT GGATATACTT 4441 GATGGAGATT TTCACACATA TTTGAGTGAT GATTCCATTG ATTCAACAGA TGATGCTGAA

4501 AAGGAAAATT TTCCCATCGA ATTTCTTAAT AGTATTACTC CTTCGGGAAT GCCGTGTCAT

4561 AAATTAAAAT TGAAAGTGGG TGCAATCATC ATGCTATTGA GAAATCTTAA TAGTAAATGG

4621 GGTCTTTGTA ATGGTACTAG ATTTATTATC AAAAGATTAC GACCTAACAT TATCGAAGCT

4681 GAAGTATTAA CAGGATCTGC AGAGGGAGAG GTTGTTCTGA TTCCAAGAAT TGATTTGTCC

4741 CCATCTGACA CTGGCCTCCC ATTTAAATTA ATTCGAAGAC AGTTTCCCGT GATGCCAGCA

4801 TTTGCGATGA CTATTAATAA ATCACAAGGA CAAACTCTAG ACAGAGTAGG AATATTCCTA

4861 CCTGAACCCG TTTTCGCACA TGGTCAGTTA TATGTTGCTT TCTCTCGAGT TCGAAGAGCA

4921 TGTGACGTTA AAGTTAAAGT TGTAAATACT TCATCACAAG GGAAATTAGT CAAGCACTCT

4981 GAAAGTGTTT TTACTCTTAA TGTGGTATAC AGGGAGATAT TAGAATAAGT TTAATCACTT

5041 TATCAGTCAT TGTTTGCATC AATGTTGTTT TTATATCATG TTTTTGTTGT TTTTATATCA

5101 TGTCTTTGTT GTTGTTATAT CATGTTGTTA TTGTTTATTT ATTAATAAAT TTATGTATTA

5161 TTTTCATATA CATTTTACTC ATTTCCTTTC ATCTCTCACA CTTCTATTAT AGAGAAAGGG

5221 CAAATAGCAA TATTAAAATA TTTCCTCTAA TTAATTCCCT TTCAATGTGC ACGAATTTCG

5281 TGCACCGGGC CACTAG (SEQ ID NO: 14652) .

[0251] Unlike other transposases, the Helitron transposase does not contain an RNase-H like catalytic domain, but instead comprises a RepHel motif made up of a replication initiator domain (Rep) and a DNA helicase domain. The Rep domain is a nuclease domain of the HUH superfamily of nucleases.

[0252] An exemplary Helitron transposase of the disclosure comprises an amino acid sequence comprising:

1 MSKEQLLIQR SSAAERCRRY RQKMSAEQRA SDLERRRRLQ QNVSEEQLLE KRRSEAEKQR

61 RHRQKMSKDQ RAFEVERRRW RRQNMSREQS STSTTNTGRN CLLSKNGVHE DAILEHSCGG

121 MTVRCEFCLS LNFSDEKPSD GKFTRCCSKG KVCPNDIHFP DYPAYLKRLM TNEDSDSKNF

181 MENIRSINSS FAFASMGANI ASPSGYGPYC FRIHGQVYHR TGTLHPSDGV SRKFAQLYIL

241 DTAEATSKRL AMPENQGCSE RLMININNLM HEINELTKSY KMLHEVEKEA QSEAAAKGIA

301 PTEVTMAIKY DRNSDPGRYN SPRVTEVAVI FRNEDGEPPF ERDLLIHCKP DPNNPNATKM

361 KQISILFPTL DAMTYPILFP HGEKGWGTDI ALRLRDNSVI DNNTRQNVRT RVTQMQYYGF

421 HLSVRDTFNP ILNAGKLTQQ FIVDSYSKME ANRINFIKAN QSKLRVEKYS GLMDYLKSRS

481 ENDNVPIGKM IILPSSFEGS PRNMQQRYQD AMAIVTKYGK PDLFITMTCN PKWADITNNL

541 QRWQKVENRP DLVARVFNIK LNALLNDICK FHLFGKVIAK IHVIEFQKRG LPHAHILLIL

601 DSESKLRSED DIDRIVKAEI PDEDQCPRLF QIVKSNMVHG PCGIQNPNSP CMENGKCSKG

661 YPKEFQNATI GNIDGYPKYK RRSGSTMSIG NKWDNTWIV PYNPYLCLKY NCHINVEVCA

721 SIKSVKYLFK YIYKGHDCAN IQISEKNIIN HDEVQDFIDS RYVSAPEAVW RLFAMRMHDQ

781 SHAITRLAIH LPNDQNLYFH TDDFAEVLDR AKRHNSTLMA WFLLNREDSD ARNYYYWEIP

841 QHYVFNNSLW TKRRKGGNKV LGRLFTVSFR EPERYYLRLL LLHVKGAISF EDLRTVGGVT

901 YDTFHEAAKH RGLLLDDTIW KDTIDDAIIL NMPKQLRQLF AYICVFGCPS AADKLWDENK

961 SHFIEDFCWK LHRREGACVN CEMHALNEIQ EVFTLHGMKC SHFKLPDYPL LMNANTCDQL

1021 YEQQQAEVLI NSLNDEQLAA FQTITSAIED QTVHPKCFFL DGPGGSGKTY LYKVLTHYIR

1081 GRGGTVLPTA STGIAANLLL GGRTFHSQYK LPIPLNETSI SRLDIKSEVA KTIKKAQLLI 1141 IDECTMASSH AINAIDRLLR EIMNLNVAFG GKVLLLGGDF RQCLSIVPHA MRSAIVQTSL 1201 KYCNVWGCFR KLSLKTNMRS EDSAYSEWLV KLGDGKLDSS FHLGMDI IEI PHEMICNGSI 1261 IEATFGNSIS IDNIKNISKR AILCPKNEHV QKLNEEILDI LDGDFHTYLS DDSIDSTDDA 1321 EKENFPIEFL NSITPSGMPC HKLKLKVGAI IMLLRNLNSK WGLCNGTRFI IKRLRPNI IE 1381 AEVLTGSAEG EWLIPRIDL SPSDTGLPFK LIRRQFPVMP AFAMTINKSQ GQTLDRVGIF 1441 LPEPVFAHGQ LYVAFSRVRR ACDVKVKW TSSQGKLVKH SESVFTLNW YREILE (SEQ ID NO: 14501 ) .

[0253] In Helitron transpositions, a hairpin close to the 3' end of the transposon functions as a terminator. However, this hairpin can be bypassed by the transposase, resulting in the transduction of flanking sequences. In addition, Helraiser transposition generates covalently closed circular intermediates. Furthermore, Helitron transpositions can lack target site duplications. In the Helraiser sequence, the transposase is flanked by left and right terminal sequences termed LTS and RTS. These sequences terminate with a conserved 5'-TC/CTAG- 3' motif. A 19 bp palindromic sequence with the potential to form the hairpin termination structure is located 11 nucleotides upstream of the RTS and consists of the sequence

GTGCACGAATTTCGTGCACCGGGCCACTAG (SEQ ID NO: 14500).

[0254] Tol2 transposons may be isolated or derived from the genome of the medaka fish, and may be similar to transposons of the hAT family. Exemplary Tol2 transposons of the disclosure are encoded by a sequence comprising about 4.7 kilobases and contain a gene encoding the Tol2 transposase, which contains four exons. An exemplary Tol2 transposase of the disclosure comprises an amino acid sequence comprising the following:

1 MEEVCDSSAA ASSTVQNQPQ DQEHPWPYLR EFFSLSGVNK DSFKMKCVLC LPLNKEISAF

61 KSSPSNLRKH IERMHPNYLK NYSKLTAQKR KIGTSTHASS SKQLKVDSVF PVKHVSPVTV

121 NKAILRYIIQ GLHPFSTVDL PSFKELISTL QPGISVITRP TLRSKIAEAA LIMKQKVTAA

181 MSEVEWIATT TDCWTARRKS FIGVTAHWIN PGSLERHSAA LACKRLMGSH TFEVLASAMN

241 DIHSEYEIRD KWCTTTDSG SNFMKAFRVF GVENNDIETE ARRCESDDTD SEGCGEGSDG

301 VEFQDASRVL DQDDGFEFQL PKHQKCACHL LNLVSSVDAQ KALSNEHYKK LYRSVFGKCQ

361 ALWNKSSRSA LAAEAVESES RLQLLRPNQT RWNSTFMAVD RILQICKEAG EGALRNICTS

421 LEVPMFNPAE MLFLTEWANT MRPVAKVLDI LQAETNTQLG WLLPSVHQLS LKLQRLHHSL

481 RYCDPLVDAL QQGIQTRFKH MFEDPEI IAA AILLPKFRTS WTNDETIIKR GMDYIRVHLE

541 PLDHKKELAN SSSDDEDFFA SLKPTTHEAS KELDGYLACV SDTRESLLTF PAICSLSIKT

601 NTPLPASAAC ERLFSTAGLL FSPKRARLDT NNFENQLLLK LNLRFYNFE (SEQ ID NO:

14502).

[0255] An exemplary Tol2 transposon of the disclosure, including inverted repeats, subterminal sequences and the Tol2 transposase, is encoded by a nucleic acid sequence comprising the following:

1 CAGAGGTGTA AAGTACTTGA GTAATTTTAC TTGATTACTG TACTTAAGTA TTATTTTTGG 61 GGATTTTTAC TTTACTTGAG TACAATTAAA AATCAATACT TTTACTTTTA CTTAATTACA

121 TTTTTTTAGA AAAAAAAGTA CTTTTTACTC CTTACAATTT TATTTACAGT CAAAAAGTAC

181 TTATTTTTTG GAGATCACTT CATTCTATTT TCCCTTGCTA TTACCAAACC AATTGAATTG

241 CGCTGATGCC CAGTTTAATT TAAATGTTAT TTATTCTGCC TATGAAAATC GTTTTCACAT

301 TATATGAAAT TGGTCAGACA TGTTCATTGG TCCTTTGGAA GTGACGTCAT GTCACATCTA

361 TTACCACAAT GCACAGCACC TTGACCTGGA AATTAGGGAA ATTATAACAG TCAATCAGTG

421 GAAGAAAATG GAGGAAGTAT GTGATTCATC AGCAGCTGCG AGCAGCACAG TCCAAAATCA

481 GCCACAGGAT CAAGAGCACC CGTGGCCGTA TCTTCGCGAA TTCTTTTCTT TAAGTGGTGT

541 AAATAAAGAT TCATTCAAGA TGAAATGTGT CCTCTGTCTC CCGCTTAATA AAGAAATATC

601 GGCCTTCAAA AGTTCGCCAT CAAACCTAAG GAAGCATATT GAGGTAAGTA CATTAAGTAT

661 TTTGTTTTAC TGATAGTTTT TTTTTTTTTT TTTTTTTTTT TTTTTGGGTG TGCATGTTTT

721 GACGTTGATG GCGCGCCTTT TATATGTGTA GTAGGCCTAT TTTCACTAAT GCATGCGATT

781 GACAATATAA GGCTCACGTA ATAAAATGCT AAAATGCATT TGTAATTGGT AACGTTAGGT

841 CCACGGGAAA TTTGGCGCCT ATTGCAGCTT TGAATAATCA TTATCATTCC GTGCTCTCAT

901 TGTGTTTGAA TTCATGCAAA ACACAAGAAA ACCAAGCGAG AAATTTTTTT CCAAACATGT

961 TGTATTGTCA AAACGGTAAC ACTTTACAAT GAGGTTGATT AGTTCATGTA TTAACTAACA

1021 TTAAATAACC ATGAGCAATA CATTTGTTAC TGTATCTGTT AATCTTTGTT AACGTTAGTT

1081 AATAGAAATA CAGATGTTCA TTGTTTGTTC ATGTTAGTTC ACAGTGCATT AACTAATGTT

1141 AACAAGATAT AAAGTATTAG TAAATGTTGA AATTAACATG TATACGTGCA GTTCATTATT

1201 AGTTCATGTT AACTAATGTA GTTAACTAAC GAACCTTATT GTAAAAGTGT TACCATCAAA

1261 ACTAATGTAA TGAAATCAAT TCACCCTGTC ATGTCAGCCT TACAGTCCTG TGTTTTTGTC

1321 AATATAATCA GAAATAAAAT TAATGTTTGA TTGTCACTAA ATGCTACTGT ATTTCTAAAA

1381 TCAACAAGTA TTTAACATTA TAAAGTGTGC AATTGGCTGC AAATGTCAGT TTTATTAAAG

1441 GGTTAGTTCA CCCAAAAATG AAAATAATGT CATTAATGAC TCGCCCTCAT GTCGTTCCAA

1501 GCCCGTAAGA CCTCCGTTCA TCTTCAGAAC ACAGTTTAAG ATATTTTAGA TTTAGTCCGA

1561 GAGCTTTCTG TGCCTCCATT GAGAATGTAT GTACGGTATA CTGTCCATGT CCAGAAAGGT

1621 AATAAAAACA TCAAAGTAGT CCATGTGACA TCAGTGGGTT AGTTAGAATT TTTTGAAGCA

1681 TCGAATACAT TTTGGTCCAA AAATAACAAA ACCTACGACT TTATTCGGCA TTGTATTCTC

1741 TTCCGGGTCT GTTGTCAATC CGCGTTCACG ACTTCGCAGT GACGCTACAA TGCTGAATAA

1801 AGTCGTAGGT TTTGTTATTT TTGGACCAAA ATGTATTTTC GATGCTTCAA ATAATTCTAC

1861 CTAACCCACT GATGTCACAT GGACTACTTT GATGTTTTTA TTACCTTTCT GGACATGGAC

1921 AGTATACCGT ACATACATTT TCAGTGGAGG GACAGAAAGC TCTCGGACTA AATCTAAAAT

1981 ATCTTAAACT GTGTTCCGAA GATGAACGGA GGTGTTACGG GCTTGGAACG ACATGAGGGT

2041 GAGTCATTAA TGACATCTTT TCATTTTTGG GTGAACTAAC CCTTTAATGC TGTAATCAGA

2101 GAGTGTATGT GTAATTGTTA CATTTATTGC ATACAATATA AATATTTATT TGTTGTTTTT

2161 ACAGAGAATG CACCCAAATT ACCTCAAAAA CTACTCTAAA TTGACAGCAC AGAAGAGAAA

2221 GATCGGGACC TCCACCCATG CTTCCAGCAG TAAGCAACTG AAAGTTGACT CAGTTTTCCC

2281 AGTCAAACAT GTGTCTCCAG TCACTGTGAA CAAAGCTATA TTAAGGTACA TCATTCAAGG

2341 ACTTCATCCT TTCAGCACTG TTGATCTGCC ATCATTTAAA GAGCTGATTA GTACACTGCA

2401 GCCTGGCATT TCTGTCATTA CAAGGCCTAC TTTACGCTCC AAGATAGCTG AAGCTGCTCT

2461 GATCATGAAA CAGAAAGTGA CTGCTGCCAT GAGTGAAGTT GAATGGATTG CAACCACAAC 2521 GGATTGTTGG ACTGCACGTA GAAAGTCATT CATTGGTGTA ACTGCTCACT GGATCAACCC

2581 TGGAAGTCTT GAAAGACATT CCGCTGCACT TGCCTGCAAA AGATTAATGG GCTCTCATAC

2641 TTTTGAGGTA CTGGCCAGTG CCATGAATGA TATCCACTCA GAGTATGAAA TACGTGACAA

2701 GGTTGTTTGC ACAACCACAG ACAGTGGTTC CAACTTTATG AAGGCTTTCA GAGTTTTTGG

2761 TGTGGAAAAC AATGATATCG AGACTGAGGC AAGAAGGTGT GAAAGTGATG ACACTGATTC

2821 TGAAGGCTGT GGTGAGGGAA GTGATGGTGT GGAATTCCAA GATGCCTCAC GAGTCCTGGA

2881 CCAAGACGAT GGCTTCGAAT TCCAGCTACC AAAACATCAA AAGTGTGCCT GTCACTTACT

2941 TAACCTAGTC TCAAGCGTTG ATGCCCAAAA AGCTCTCTCA AATGAACACT ACAAGAAACT

3001 CTACAGATCT GTCTTTGGCA AATGCCAAGC TTTATGGAAT AAAAGCAGCC GATCGGCTCT

3061 AGCAGCTGAA GCTGTTGAAT CAGAAAGCCG GCTTCAGCTT TTAAGGCCAA ACCAAACGCG

3121 GTGGAATTCA ACTTTTATGG CTGTTGACAG AATTCTTCAA ATTTGCAAAG AAGCAGGAGA

3181 AGGCGCACTT CGGAATATAT GCACCTCTCT TGAGGTTCCA ATGTAAGTGT TTTTCCCCTC

3241 TATCGATGTA AACAAATGTG GGTTGTTTTT GTTTAATACT CTTTGATTAT GCTGATTTCT

3301 CCTGTAGGTT TAATCCAGCA GAAATGCTGT TCTTGACAGA GTGGGCCAAC ACAATGCGTC

3361 CAGTTGCAAA AGTACTCGAC ATCTTGCAAG CGGAAACGAA TACACAGCTG GGGTGGCTGC

3421 TGCCTAGTGT CCATCAGTTA AGCTTGAAAC TTCAGCGACT CCACCATTCT CTCAGGTACT

3481 GTGACCCACT TGTGGATGCC CTACAACAAG GAATCCAAAC ACGATTCAAG CATATGTTTG

3541 AAGATCCTGA GATCATAGCA GCTGCCATCC TTCTCCCTAA ATTTCGGACC TCTTGGACAA

3601 ATGATGAAAC CATCATAAAA CGAGGTAAAT GAATGCAAGC AACATACACT TGACGAATTC

3661 TAATCTGGGC AACCTTTGAG CCATACCAAA ATTATTCTTT TATTTATTTA TTTTTGCACT

3721 TTTTAGGAAT GTTATATCCC ATCTTTGGCT GTGATCTCAA TATGAATATT GATGTAAAGT

3781 ATTCTTGCAG CAGGTTGTAG TTATCCCTCA GTGTTTCTTG AAACCAAACT CATATGTATC

3841 ATATGTGGTT TGGAAATGCA GTTAGATTTT ATGCTAAAAT AAGGGATTTG CATGATTTTA

3901 GATGTAGATG ACTGCACGTA AATGTAGTTA ATGACAAAAT CCATAAAATT TGTTCCCAGT

3961 CAGAAGCCCC TCAACCAAAC TTTTCTTTGT GTCTGCTCAC TGTGCTTGTA GGCATGGACT

4021 ACATCAGAGT GCATCTGGAG CCTTTGGACC ACAAGAAGGA ATTGGCCAAC AGTTCATCTG

4081 ATGATGAAGA TTTTTTCGCT TCTTTGAAAC CGACAACACA TGAAGCCAGC AAAGAGTTGG

4141 ATGGATATCT GGCCTGTGTT TCAGACACCA GGGAGTCTCT GCTCACGTTT CCTGCTATTT

4201 GCAGCCTCTC TATCAAGACT AATACACCTC TTCCCGCATC GGCTGCCTGT GAGAGGCTTT

4261 TCAGCACTGC AGGATTGCTT TTCAGCCCCA AAAGAGCTAG GCTTGACACT AACAATTTTG

4321 AGAATCAGCT TCTACTGAAG TTAAATCTGA GGTTTTACAA CTTTGAGTAG CGTGTACTGG

4381 CATTAGATTG TCTGTCTTAT AGTTTGATAA TTAAATACAA ACAGTTCTAA AGCAGGATAA

4441 AACCTTGTAT GCATTTCATT TAATGTTTTT TGAGATTAAA AGCTTAAACA AGAATCTCTA

4501 GTTTTCTTTC TTGCTTTTAC TTTTACTTCC TTAATACTCA AGTACAATTT TAATGGAGTA

4561 CTTTTTTACT TTTACTCAAG TAAGATTCTA GCCAGATACT TTTACTTTTA ATTGAGTAAA

4621 ATTTTCCCTA AGTACTTGTA CTTTCACTTG AGTAAAATTT TTGAGTACTT TTTACACCTC

4681 TG (SEQ ID NO: 14653).

[0256] Exemplary transposon/transposase systems of the disclosure include, but are not limited to, piggyBac and piggyBac-like transposons and transposases. [0257] PiggyBac and piggyBac-like transposases recognizes transposon-specific inverted terminal repeat sequences (ITRs) on the ends of the transposon, and moves the contents between the ITRs into TTAA or TTAT chromosomal sites. The piggyBac or piggyBac-like transposon system has no payload limit for the genes of interest that can be included between the ITRs.

[0258] In certain embodiments, and, in particular, those embodiments wherein the transposon is a piggyBac transposon, the transposase is a piggyBac™, Super piggyBac™ (SPB) transposase. In certain embodiments, and, in particular, those embodiments wherein the transposase is a piggyBac™, Super piggyBac™ (SPB), the sequence encoding the transposase is an mRNA sequence.

[0259] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme.

[0260] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or a piggyBac-like transposase enzyme. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG 61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG 121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF 181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV 241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKILMMCD 301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ 361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC 421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN 481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO: 14487) .

[0261] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at one or more of positions 30, 165, 282, or 538 of the sequence:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEI SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG 61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG 121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTGATFRD TNEDEIYAFF 181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV 241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RMYIPNKPSK YGIKIL MCD 301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ 361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC 421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN 481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPNEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO: 14487) .

[0262] In certain embodiments, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at two or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at three or more of positions 30, 165, 282, or 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme that comprises or consists of an amino acid sequence having an amino acid substitution at each of the following positions 30, 165, 282, and 538 of the sequence of SEQ ID NO: 14487. In certain embodiments, the amino acid substitution at position 30 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 165 of the sequence of SEQ ID NO: 14487 is a substitution of a serine (S) for a glycine (G). In certain embodiments, the amino acid substitution at position 282 of the sequence of SEQ ID NO: 14487 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 538 of the sequence of SEQ ID NO: 14487 is a substitution of a lysine (K) for an asparagine (N).

[0263] In certain embodiments of the methods of the disclosure, the transposase enzyme is a Super piggyBac™ (SPB) or piggyBac-like transposase enzyme. In certain embodiments, the Super piggyBac™ (SPB) or piggyBac-like transposase enzyme of the disclosure may comprise or consist of the amino acid sequence of the sequence of SEQ ID NO: 14487 wherein the amino acid substitution at position 30 is a substitution of a valine (V) for an isoleucine (I), the amino acid substitution at position 165 is a substitution of a serine (S) for a glycine (G), the amino acid substitution at position 282 is a substitution of a valine (V) for a methionine (M), and the amino acid substitution at position 538 is a substitution of a lysine (K) for an asparagine (N). In certain embodiments, the Super piggyBac™ (SPB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG 61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG 121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF 181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV 241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD 301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ 361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC 421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN 481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ ID NO: 14484) .

[0264] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™, Super piggyBac™ or piggyBac-like transposase enzyme may further comprise an amino acid substitution at one or more of positions 3, 46, 82, 103, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 258, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 486, 503, 552, 570 and 591 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™, Super piggyBac™ or piggyBac-like transposase enzyme may further comprise an amino acid substitution at one or more of positions 46, 119, 125, 177, 180, 185, 187, 200, 207, 209, 226, 235, 240, 241, 243, 296, 298, 311, 315, 319, 327, 328, 340, 421, 436, 456, 470, 485, 503, 552 and 570. In certain embodiments, the amino acid substitution at position 3 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for a serine (S). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an alanine (A). In certain embodiments, the amino acid substitution at position 46 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 82 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for an isoleucine (I). In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 119 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for an arginine (R). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) a cysteine (C). In certain embodiments, the amino acid substitution at position 125 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 177 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 180 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 185 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 187 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for an alanine (A). In certain embodiments, the amino acid substitution at position 200 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 207 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a valine (V). In certain embodiments, the amino acid substitution at position 209 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a valine (V). In certain embodiments, the amino acid substitution at position 226 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a methionine (M). In certain embodiments, the amino acid substitution at position 235 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a leucine (L). In certain embodiments, the amino acid substitution at position 240 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 241 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a phenylalanine (F). In certain embodiments, the amino acid substitution at position 243 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a proline (P). In certain embodiments, the amino acid substitution at position 258 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tryptophan (W) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a leucine (L). In certain embodiments, the amino acid substitution at position 296 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a methionine (M). In certain embodiments, the amino acid substitution at position 298 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a proline (P). In certain embodiments, the amino acid substitution at position 311 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine for a proline (P). In certain embodiments, the amino acid substitution at position 315 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for an arginine (R).In certain embodiments, the amino acid substitution at position 319 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a threonine (T). In certain embodiments, the amino acid substitution at position 327 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 328 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a tyrosine (Y). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a cysteine (C). In certain embodiments, the amino acid substitution at position 340 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a cysteine (C). In certain embodiments, the amino acid substitution at position 421 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a histidine (H) for the aspartic acid (D). In certain embodiments, the amino acid substitution at position 436 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a valine (V). In certain embodiments, the amino acid substitution at position 456 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a tyrosine (Y) for a methionine (M). In certain embodiments, the amino acid substitution at position 470 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a

phenylalanine (F) for a leucine (L). In certain embodiments, the amino acid substitution at position 485 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a serine (S). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a leucine (L) for a methionine (M). In certain embodiments, the amino acid substitution at position 503 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an isoleucine (I) for a methionine (M). In certain embodiments, the amino acid substitution at position 552 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a lysine (K) for a valine (V). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a threonine (T) for an alanine (A). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a glutamine (Q). In certain embodiments, the amino acid substitution at position 591 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an arginine (R) for a glutamine (Q).

[0265] In certain embodiments of the methods of the disclosure, including those

embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or piggyBac-like transposase enzyme or may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at one or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments of the methods of the disclosure, including those embodiments wherein the transposase comprises the above- described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or piggyBac-like transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at two, three, four, five, six or more of positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, including those embodiments wherein the transposase comprises the above-described mutations at positions 30, 165, 282 and/or 538, the piggyBac™ or piggyBac-like transposase enzyme may comprise or the Super piggyBac™ transposase enzyme may further comprise an amino acid substitution at positions 103, 194, 372, 375, 450, 509 and 570 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the amino acid substitution at position 103 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a proline (P) for a serine (S). In certain embodiments, the amino acid substitution at position 194 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a valine (V) for a methionine (M). In certain embodiments, the amino acid substitution at position 372 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for an arginine (R). In certain embodiments, the amino acid substitution at position 375 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an alanine (A) for a lysine (K). In certain embodiments, the amino acid substitution at position 450 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of an asparagine (N) for an aspartic acid (D). In certain embodiments, the amino acid substitution at position 509 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a glycine (G) for a serine (S). In certain embodiments, the amino acid substitution at position 570 of SEQ ID NO: 14487 or SEQ ID NO: 14484 is a substitution of a serine (S) for an asparagine (N). In certain embodiments, the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487. In certain embodiments, including those embodiments wherein the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, the piggyBac™ or piggyBac-like transposase enzyme may further comprise an amino acid substitution at positions 372, 375 and 450 of the sequence of SEQ ID NO: 14487 or SEQ ID NO: 14484. In certain embodiments, the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, and a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487. In certain embodiments, the piggyBac™ or piggyBac-like transposase enzyme may comprise a substitution of a valine (V) for a methionine (M) at position 194 of SEQ ID NO: 14487, a substitution of an alanine (A) for an arginine (R) at position 372 of SEQ ID NO: 14487, a substitution of an alanine (A) for a lysine (K) at position 375 of SEQ ID NO: 14487 and a substitution of an asparagine (N) for an aspartic acid (D) at position 450 of SEQ ID NO: 14487.

[0266] In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from an insect. In certain embodiments, the insect is Trichoplusia ni (GenBank Accession No. AAA87375; SEQ ID NO: 14666), Argyrogramma agnata

(GenBank Accession No. GU477713; SEQ ID NO: 14534, SEQ ID NO: 14667), Anopheles gambiae (GenBank Accession No. XP_312615 (SEQ ID NO: 14668); GenBank Accession No. XP_320414 (SEQ ID NO: 14669); GenBank Accession No. XP_310729 (SEQ ID NO: 14670)), Aphis gossypii (GenBank Accession No. GU329918; SEQ ID NO: 14671, SEQ ID NO: 14672), Acyrthosiphon pisum (GenBank Accession No. XP_001948139; SEQ ID NO: 14673), Agrotis ipsilon (GenBank Accession No. GU477714; SEQ ID NO: 14537, SEQ ID NO: 14674), Bombyx mori (GenBank Accession No. BAD11135; SEQ ID NO: 14505), Chilo suppressalis (GenBank Accession No. JX294476; SEQ ID NO: 14675, SEQ ID NO: 14676), Drosophila melanogaster (GenBank Accession No. AAL39784; SEQ ID NO: 14677), Helicoverpa armigera (GenBank Accession No. ABS18391 ; SEQ ID NO: 14525), Heliothis virescens (GenBank Accession No. ABD76335; SEQ ID NO: 14678), Macdunnoughia crassisigna (GenBank Accession No. EU287451; SEQ ID NO: 14679, SEQ ID NO: 14680), Pectinophora gossypiella (GenBank Accession No. GU270322; SEQ ID NO: 14530, SEQ ID NO: 14681), Tribolium castaneum (GenBank Accession No. XP_001814566; SEQ ID NO: 14682), Ctenoplusia agnata (also called Argyrogramma agnata), Messour bouvieri,

Megachile rotundata, Bombus impatiens , Mamestra brassicae, Mayetiola destructor or Apis mellifera.

[0267] In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from an insect. In certain embodiments, the insect is Trichoplusia ni (AAA87375).

[0268] In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from an insect. In certain embodiments, the insect is Bombyx mori (BAD11135).

[0269] In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from a crustacean. In certain embodiments, the crustacean is Daphnia pulicaria (AAM76342, SEQ ID NO: 14683).

[0270] In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from a vertebrate. In certain embodiments, the vertebrate is Xenopus tropicalis (GenBank Accession No. BAF82026; SEQ ID NO: 14518), Homo sapiens (GenBank Accession No. NP_689808; SEQ ID NO: 14684), Mus musculus (GenBank Accession No. NP_741958; SEQ ID NO: 14685), Macaca fascicularis (GenBank Accession No. AB179012; SEQ ID NO: 14686, SEQ ID NO: 14687), Rattus norvegicus (GenBank Accession No. XP_220453; SEQ ID NO: 14688) or Myotis lucifugus.

[0271] In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from a urochordate. In certain embodiments, the urochordate is Ciona intestinalis (GenBank Accession No. XP_002123602; SEQ ID NO: 14689).

[0272] In certain embodiments, the piggyBac or piggyBac-like transposase inserts a transposon at the sequence 5'-TTAT-3' within a chromosomal site (a TTAT target sequence).

[0273] In certain embodiments, the piggyBac or piggyBac-like transposase inserts a transposon at the sequence 5'-TTAA-3' within a chromosomal site (a TTAA target sequence). [0274] In certain embodiments, the target sequence of the piggyBac or piggyBac-like transposon comprises or consists of 5 '-CTAA-3', 5 '-TTAG-3', 5 '-ATAA-3 ', 5'-TCAA-3', 5'AGTT-3', 5 '-ATTA-3', 5 '-GTTA-3 ', 5'-TTGA-3', 5'-TTTA-3 ', 5'-TTAC-3 ', 5'-ACTA- 3', 5 '-AGGG-3', 5 '-CTAG-3', 5'-TGAA-3', 5 '-AGGT-3', 5 '-ATCA-3', 5 '-CTCC-3', 5 '- TAAA-3', 5'-TCTC-3 ', 5 'TGAA-3', 5 '-AAAT-3', 5 '-AATC-3', 5 '-ACAA-3 ', 5'-ACAT-3 ', 5'-ACTC-3', 5 '-AGTG-3 ', 5 '-ATAG-3 ', 5'-CAAA-3', 5'-CACA-3', 5 '-CATA-3 ', 5'- CCAG-3', 5'-CCCA-3 ', 5'-CGTA-3', 5 '-GTCC-3 ', 5'-TAAG-3 ', 5'-TCTA-3', 5'-TGAG-3', 5'-TGTT-3 ', 5'-TTCA-3 '5 '-TTCT-3 ' and 5'-TTTT-3 ' .

[0275] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Bombyx mori. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181 FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FDWNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELSANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRANKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KHSCNACAKP ICMEHAKFLC

601 ENCAELDSSL (SEQ ID NO: 14504) .

[0276] The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181 FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FYWNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601 ENCAELDSSL (SEQ ID NO: 14505) .

[0277] In certain embodiments, the piggyBac or piggyBac-like transposase is fused to a nuclear localization signal. In certain embodiments, the amino acid sequence of the piggyBac or piggyBac-like transposase fused to a nuclear localization signal is encoded by a polynucleotide sequence comprising:

1 atggcaccca aaaagaaacg taaagtgatg gacattgaaa gacaggaaga aagaatcagg

61 gcgatgctcg aagaagaact gagcgactac tccgacgaat cgtcatcaga ggatgaaacc

121 gaccactgta gcgagcatga ggttaactac gacaccgagg aggagagaat cgactctgtg

181 gatgtgccct ccaactcacg ccaagaagag gccaatgcaa ttatcgcaaa cgaatcggac

241 agcgatccag acgatgatct gccactgtcc ctcgtgcgcc agcgggccag cgcttcgaga

301 caagtgtcag gtccattcta cacttcgaag gacggcacta agtggtacaa gaattgccag

361 cgacctaacg tcagactccg ctccgagaat atcgtgaccg aacaggctca ggtcaagaat

421 atcgcccgcg acgcctcgac tgagtacgag tgttggaata tcttcgtgac ttcggacatg

481 ctgcaagaaa ttctgacgca caccaacagc tcgattaggc atcgccagac caagactgca

541 gcggagaact catcggccga aacctccttc tatatgcaag agactactct gtgcgaactg

601 aaggcgctga ttgcactgct gtacttggcc ggcctcatca aatcaaatag gcagagcctc

661 aaagatctct ggagaacgga tggaactgga gtggatatct ttcggacgac tatgagcttg

721 cagcggttcc agtttctgca aaacaatatc agattcgacg acaagtccac ccgggacgaa

781 aggaaacaga ctgacaacat ggctgcgttc cggtcaatat tcgatcagtt tgtgcagtgc

841 tgccaaaacg cttatagccc atcggaattc ctgaccatcg acgaaatgct tctctccttc

901 cgggggcgct gcctgttccg agtgtacatc ccgaacaagc cggctaaata cggaatcaaa

961 atcctggccc tggtggacgc caagaatttc tacgtcgtga atctcgaagt gtacgcagga

1021 aagcaaccgt cgggaccgta cgctgtttcg aaccgcccgt ttgaagtcgt cgagcggctt

1081 attcagccgg tggccagatc ccaccgcaat gttaccttcg acaattggtt caccggctac

1141 gagctgatgc ttcaccttct gaacgagtac cggctcacta gcgtggggac tgtcaggaag

1201 aacaagcggc agatcccaga atccttcatc cgcaccgacc gccagcctaa ctcgtccgtg

1261 ttcggatttc aaaaggatat cacgcttgtc tcgtacgccc ccaagaaaaa caaggtcgtg

1321 gtcgtgatga gcaccatgca tcacgacaac agcatcgacg agtcaaccgg agaaaagcaa

1381 aagcccgaga tgatcacctt ctacaattca actaaggccg gcgtcgacgt cgtggatgaa

1441 ctgtgcgcga actataacgt gtcccggaac tctaagcggt ggcctatgac tctcttctac

1501 ggagtgctga atatggccgc aatcaacgcg tgcatcatct accgcaccaa caagaacgtg

1561 accatcaagc gcaccgagtt catcagatcg ctgggtttga gcatgatcta cgagcacctc

1621 cattcacgga acaagaagaa gaatatccct acttacctga ggcagcgtat cgagaagcag

1681 ttgggagaac caagcccgcg ccacgtgaac gtgccggggc gctacgtgcg gtgccaagat

1741 tgcccgtaca aaaaggaccg caaaaccaaa agatcgtgta acgcgtgcgc caaacctatc

1801 tgcatggagc atgccaaatt tctgtgtgaa aattgtgctg aactcgattc ctccctg

(SEQ ID NO: 14629) .

[0278] In certain embodiments, the piggyBac or piggyBac-like transposase is hyperactive. A hyperactive piggyBac or piggyBac-like transposase is a transposase that is more active than the naturally occurring variant from which it is derived. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase enzyme is isolated or derived from Bombyx mori. In certain embodiments, the piggyBac or piggyBac-like transposase is a hyperactive variant of SEQ ID NO: 14505. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE 61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQMSGPHYTS KDGTKWYKNC QRPNVRLRSE 121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSASTS 181 FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN 241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY 301 IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR 361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL 421 VSYAPKKNKV VWMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR 481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI 541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC 601 ENCAELDSHL (SEQ ID NO 14576) . [0279] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14576. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSAETS

181 FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLLNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FYVHNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YEVMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601 ENCAHLDS (SEQ ID NO: 14630) .

[0280] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSASTS

181 FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLLNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIAM QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601 ENCAELDSSL (SEQ ID NO: 14631) .

[0281] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSAETS

181 FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLLNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKTQIPENF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELQANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601 ENCAELDSSL (SEQ ID NO: 14632) .

[0282] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRWRQTKT AAENSSAETS

181 FYMQETTLCE LKALIGLLYI AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FYVKNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601 ENCAELDSSL (SEQ ID NO: 14633) . [0283] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181 FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFQFLQNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN DYWNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL

421 VSYAPKKNKV WVMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI

541 PTYLRQRIEK QLGEPSSRHV NVKGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC

601 ENCAELDSSL (SEQ ID NO: 14634) .

[0284] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase is more active than the transposase of SEQ ID NO: 14505. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase is at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% or any percentage in between identical to SEQ ID NO: 14505.

[0285] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution at a position selected from 92, 93, 96, 97, 165, 178, 189, 196, 200, 201, 211, 215, 235, 238, 246, 253, 258, 261, 263, 271, 303, 321, 324, 330, 373, 389, 399, 402, 403, 404, 448, 473, 484, 507,5 23, 527, 528, 543, 549, 550, 557,6 01, 605, 607, 609, 610 or a combination thereof (relative to SEQ ID NO: 14505). In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution of Q92A, V93L, V93M, P96G, F97H, F97C, H165E, H165W, E178S, E178H, C189P, A196G, L200I, A201Q, L211A, W215Y, G219S, Q235Y, Q235G, Q238L, K246I, K253V, M258V, F261L, S263K, C271S, N303R, F321W, F321D, V324K, V324H, A330V, L373C, L373V, V389L, S399N, R402K, T403L, D404Q, D404S, D404M, N441R, G448W, E449A, V469T, C473Q, R484K T507C, G523A, I527M, Y528K Y543I, E549A, K550M, P557S, E601V, E605H, E605W, D607H, S609H, L610I or any combination thereof. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution of Q92A, V93L, V93M, P96G, F97H, F97C, H165E, H165W, E178S, E178H, C189P, A196G, L200I, A201Q, L211A, W215Y, G219S, Q235Y, Q235G, Q238L, K246I, K253V, M258V, F261L, S263K, C271S, N303R, F321W, F321D, V324K, V324H, A330V, L373C, L373V, V389L, S399N, R402K, T403L, D404Q, D404S, D404M, N441R, G448W, E449A, V469T, C473Q, R484K T507C, G523A, I527M, Y528K Y543I, E549A, K550M, P557S, E601V, E605H, E605W, D607H, S609H and L610I. [0286] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of E4X, A12X, M13X,L14X, E15X, D20X, E24X, S25X, S26X, S27X, D32X, H33X, E36X, E44X, E45X, E46X, I48X, D49X, R58X, A62X, N63X, A64X, I65X, I66X, N68X, E69X, D71X, S72X, D76X, P79X, R84X, Q85X, A87X, S88X, Q92X, V93X, S94X, G95X, P96X, F97X, Y98X, T99X, I145X, S 149X, D150X, L152X, E154X, T157X, N160X, S161X, S162X, H165X, R166X, T168X, K169X, T170X, A171X, E173X, S175X, S 176X, E178X, T179X, M183X, Q184X, T186X, T187X, L188X, C189X, L194X, I195X, A196X, L198X, L200X, A201X, L203X, I204X, K205X, A206X, N207X, Q209X, S210X, L211X, K212X, D213X, L214X, W215X, R216X, T217X, G219X, V222X, D223X, I224X, T227X, M229X, Q235X, L237X, Q238X, N239X, N240X, P302X, N303X, P305X, A306X, K307X, Y308X, I310X, K311X, I312X, L313X, A314X, L315X, V316X,D317X, A318X, K319X, N320X, F321X, Y322X, V323X, V324X, L326X, E327X, V328X, A330X, Q333X, P334X, S335X, G336X, P337X, A339X, V340X, S341X, N342X, R343X, P344X, F345X, E346X, V347X, E349X, I352X, Q353X, V355X, A356X, R357X, N361X, D365X, W367X, T369X, G370X, L373X, M374X, L375X, H376X, N379X, E380X, R382X, V386X, V389X, N392X, R394X, Q395X, S399X, F400X, 140 IX, R402XT403X, D404X, R405X, Q406X, P407X, N408X, S409X, S410X, V411X, F412X, F414X, Q415X, I418X, T419X, L420X, N428XV432X, M434X, D440X, N441X, S442X, I443X, D444X, E445X, G448X, E449X, Q451X, K452X, M455X, I456X, T457X, F458X, S461X, A464X, V466X, Q468X, V469X, E471X, L472X, C473X, A474X, K483X, W485X, T488X, L489X, Y491X, G492X, V493X, M496X, I499X, C502X, I503X, T507X, K509X, N510X, V511X, T512X, I513X, R515X, E517X, S521X, G523X, L524X, S525X, I527X, Y528X, E529X, H532X, S533X, N535X, K536X, K537X, N539X, I540X, T542X, Y543X, Q546X, E549X, K550X, Q551X, G553X, E554X, P555X, S556X, P557X, R558X, H559X, V560X, N561X, V562X, P563X, G564X, R565X, Y566X, V567X, Q570X, D571X, P573X, Y574X, K576X, K581X, S583X, A586X, A588X, E594X, F598X, L599X, E601X, N602X, C603X, A604X, E605X, L606X, D607X, S608X, S609X or L610X (relative to SEQ ID NO: 14505). A list of hyperactive amino acid substitutions can be found in US patent No. 10,041,077, the contents of which are incorporated herein by reference in their entirety.

[0287] In certain embodiments, the piggyBac or piggyBac-like transposase is integration deficient. In certain embodiments, an integration deficient piggyBac or piggyBac-like transposase is a transposase that can excise its corresponding transposon, but that integrates the excised transposon at a lower frequency than a corresponding wild type transposase. In certain embodiments, the piggyBac or piggyBac-like transposase is an integration deficient variant of SEQ ID NO: 14505.

[0288] In certain embodiments, the excision competent, integration deficient piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of R9X, A12X, M13X, D20X, Y21K, D23X, E24X, S25X, S26X, S27X, E28X, E30X, D32X, H33X, E36X, H37X, A39X, Y41X, D42X, T43X, E44X, E45X, E46X, R47X, D49X, S50X, S55X, A62X, N63X, A64X, I66X, A67X, N68X, E69X, D70X, D71X, S72X, D73X, P74X, D75X, D76X, D77X,I78X, S81X,V83X, R84X, Q85X, A87X, S88X,

A89X,S90X,R91X, Q92X, V93X, S94X, G95X, P96X, F97X, Y98X, T99X, W012X, G103X, Y107X, K108X, L117X, I122X, Q128X, I312X, D135X, S137X, E139X, Y140X, I145X, S149X, D150X, Q153X, E154X, T157X, S161X, S162X, R164X, H165X, R166X, Q167X, T168X, K169X, T170X, A171X, A172X, E173X, R174X, S175X, S176X, A177X, E178X, T179X, S180X,Y182X, Q184X, E185X, T187X, L188X, C189X, L194X, I195X, A196X, L198X, L200X, A201X, L203X, I204X, K205X, N207X, Q209X, L211X, D213X, L214X, W215X, R216X, T217X, G219X, T220X, V222X, D223X, I224X, T227X, T228X, F234X, Q235X, L237X, Q238X, N239X, N240X, N303X, K304X, I310X, I312X, L313X, A314X, L315X, V316X,D317X, A318X, K319X, N320X, F321X, Y322X, V323X, V324X, N325X, L326X, E327X, V328X, A330X, G331X, K332X, Q333X, S335X, P337X, P344X, F345X, E349X, H359X, N361X, V362X, D365X, F368X, Y371X, E372X, L373X, H376X, E380X, R382X, R382X, V386X, G387X, T388X, V389X, K391X, N392X, R394X, Q395X, E398X, S399X, F400X, I401X, R402XT403X, D404X, R405X, Q406X, P407X, N408X, S409X, S410X, Q415X,K416X, A424X, K426X, N428X, V430X, V432X, V433X, M434X, D436X, D440X, N441X, S442X, I443X, D444X, E445X, S446X, T447X, G448X, E449X, K450X, Q451X, E454X, M455X, I456X, T457X, F458X, S461X, A464X, V466X, Q468X, V469X, C473X, A474X, N475X, N477X, K483X, R484X, P486X, T488X, L489X, G492X, V493X, M496X, I499X, I503X, Y505X, T507X, N510X, V511X, T512X, I513X, K514X, T516X, E517X, S521X, G523X, L524X, S525X, I527X, Y528X, L531X, H532X, S533X, N535X, I540X, T542X, Y543X, R545X, Q546X, E549X, L552X, G553X, E554X, P555X, S556X, P557X, R558X, H559X, V560X, N561X, V562X, P563X, G564X, V567X, Q570X, D571X, P573X, Y574X, K575X, K576X, N585X, A586X, M593X, K596X, E601X, N602X, A604X, E605X, L606X, D607X, S608X, S609X or L610X (relative to SEQ ID NO: 14505). A list of integration deficient amino acid substitutions can be found in US patent No.

10,041,077, the contents of which are incorporated by reference in their entirety.

[0289] In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181 FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRKDGT GVDIFRTTMS LQRFQFLLNN

241 IRFDDISTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN FYWNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL 421 VSYAPKKNKV VWMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKKWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLS MYEH LHSRNKKKNI 541 PTYLRQRIEK QLGEPVPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC 601 ENCAELDSSL (SEQ ID NO : 14606) .

In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE 61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE 121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS 181 FYMQETTLCE LKALIGLLYL AGLIKSNRQS LKDLWRTDGT GVDIFRTTMS LQRFYFLQNN 241 IRFDDKSTLD ERKQTDNMAA FRSIFDQFVQ SCQNAYSPSE FLTIDEMLLS FRGRCLFRVY 301 IPNKPAKYGI KILALVDAKN FYWNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR 361 NVTFDNWFTG YELMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL 421 VSYAPKKNKV VWMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR 481 NSKRWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIYEH LHSRNKKKNI 541 PTYLRQRIEK QLGEPSPRHV NYPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC 601 VNCAELDSSL (SEQ ID NO: 14607) .

In certain embodiments, the piggyBac or piggyBac-like transposase that is is integration deficient comprises a sequence of:

1 MDIERQEERI RAMLEEELSD YSDESSSEDE TDHCSEHEVN YDTEEERIDS VDVPSNSRQE

61 EANAI IANES DSDPDDDLPL SLVRQRASAS RQVSGPFYTS KDGTKWYKNC QRPNVRLRSE

121 NIVTEQAQVK NIARDASTEY ECWNIFVTSD MLQEILTHTN SSIRHRQTKT AAENSSAETS

181 FYMQETTLCE LKALIALLYL AGLIKSNRQS LKDLWRKDGT GVDIFRTTMS LQRFQFLLNN

241 IRFDDKSTRD ERKQTDNMAA FRSIFDQFVQ CCQNAYSPSE FLTIDEMLLS FRGRCLFRVY

301 IPNKPAKYGI KILALVDAKN DYWNLEVYA GKQPSGPYAV SNRPFEWER LIQPVARSHR

361 NVTFDNWFTG YECMLHLLNE YRLTSVGTVR KNKRQIPESF IRTDRQPNSS VFGFQKDITL 421 VSYAPKKNKV VWMSTMHHD NSIDESTGEK QKPEMITFYN STKAGVDWD ELCANYNVSR

481 NSKKWPMTLF YGVLNMAAIN ACIIYRTNKN VTIKRTEFIR SLGLSMIKEH LHSRNKKKNI 541 PTYLRQRIEK QLGEPSPRHV NVPGRYVRCQ DCPYKKDRKT KRSCNACAKP ICMEHAKFLC 601 ENCAELDSSL (SEQ ID NO: 14608) .

In certain embodiments, the integration deficient transposase comprises a sequence that is at least 90% identical to SEQ ID NO: 14608.

[0290] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Bombyx mori. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttatcccggc gagcatgagg cagggtatct cataccctgg taaaatttta aagttgtgta 61 ttttataaaa ttttcgtctg acaacactag cgcgctcagt agctggaggc aggagcgtgc 121 gggaggggat agtggcgtga tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc 181 aaacctgttt cgggtatgtt ataccctgcc tcattgttga cgtatttttt ttatgtaatt 241 tttccgatta ttaatttcaa ctgttttatt ggtattttta tgttatccat tgttcttttt 301 ttatgattta ctgtatcggt tgtctttcgt tcctttagtt gagttttttt ttattatttt 361 cagtttttga tcaaa (SEQ ID NO: 14506) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct 61 ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt 121 gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa 181 taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataaac cattatttta 241 caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa 301 atatgggtat gtcataccct gccacattct tgatgtaact ttttttcacc tcatgctcgc 361 cgggttat (SEQ ID NO: 14507) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttatcccggc gagcatgagg cagggtatct cataccctgg taaaatttta aagttgtgta 61 ttttataaaa ttttcgtctg acaacactag cgcgctcagt agctggaggc aggagcgtgc 121 gggaggggat agtggcgtga tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc 181 aaacctgttt cgggtatgtt ataccctgcc teat (SEQ ID NO: 14508).

In certain embodiments, the piggyBac™ (PB) or piggyBac-like transposon comprises a sequence of:

1 taaataataa taatttcata attaaaaact tctttcattg aatgecatta aataaaccat 61 tattttacaa aataagatca acataattga gtaaataata ataagaacaa tattatagta 121 caacaaaata tgggtatgtc ataccctgcc acattcttga tgtaactttt tttcacctca 181 tgctcgccgg gttat (SEQ ID NO: 14509) .

[0291] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left sequence corresponding to SEQ ID NO: 14506 and a right sequence corresponding to SEQ ID NO: 14507. In certain embodiments, one piggyBac or piggyBac-like transposon end is at least 85%, at least 90%, at least 95%, at least 98%, at least 99% identical or any percentage in between identical to SEQ ID NO: 14506 and the other piggyBac or piggyBac-like transposon end is at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or any percentage in between identical to SEQ ID NO: 14507. In certain embodiments, the piggyBac or piggyBac- like transposon comprises SEQ ID NO: 14506 and SEQ ID NO: 14507 or SEQ ID NO: 14509. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14508 and SEQ ID NO: 14507 or SEQ ID NO: 14509. In certain embodiments, the left and right transposon ends share a 16 bp repeat sequence at their ends of

CCCGGCGAGCATGAGG (SEQ ID NO: 14510) immediately adjacent to the 5'-TTAT-3 target insertion site, which is inverted in the orientation in the two ends. In certain embodiments, left transposon end begins with a sequence comprising 5'- TTATCCCGGCGAGCATGAGG-3 (SEQ ID NO: 14511), and the right transposon ends with a sequence comprising the reverse complement of this sequence: 5'-

CCTCATGCTCGCCGGGTTAT-3' (SEQ ID NO: 14512).

[0292] In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides of SEQ ID NO: 14506 or SEQ ID NO: 14508. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides of SEQ ID NO: 14507 or SEQ ID NO: 14509. In certain embodiments, the piggyBac or piggyBac- like transposon comprises one end with at least 90% identity to SEQ ID NO: 14506 or SEQ ID NO: 14508. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14507 or SEQ ID NO: 14509.

[0293] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaacccggc gagcatgagg cagggtatct cataccctgg taaaatttta aagttgtgta 61 ttttataaaa ttttcgtctg acaacactag cgcgctcagt agctggaggc aggagcgtgc 121 gggaggggat agtggcgtga tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc 181 aaacctgttt cgggtatgtt ataccctgcc tcattgttga cgtatttttt ttatgtaatt 241 tttccgatta ttaatttcaa ctgttttatt ggtattttta tgttatccat tgttcttttt 301 ttatgattta ctgtatcggt tgtctttcgt tcctttagtt gagttttttt ttattatttt 361 cagtttttga tcaaa (SEQ ID NO: 14515) .

[0294] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct

61 ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt

121 gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa

181 taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataatt cattatttta

241 caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa

301 atatgggtat gtcataccct tttttttttt tttttttttt ttttttcggg tagagggccg

361 aacctcctac gaggtccccg cgcaaaaggg gcgcgcgggg tatgtgagac tcaacgatct

421 gcatggtgtt gtgagcagac cgcgggccca aggattttag agcccaccca ctaaacgact

481 cctctgcact cttacacccg acgtccgatc ccctccgagg tcagaacccg gatgaggtag

541 gggggctacc gcggtcaaca ctacaaccag acggcgcggc tcaccccaag gacgcccagc

601 cgacggagcc ttcgaggcga atcgaaggct ctgaaacgtc ggccgtctcg gtacggcagc

661 ccgtcgggcc gcccagacgg tgccgctggt gtcccggaat accccgctgg accagaacca

721 gcctgccggg tcgggacgcg atacaccgtc gaccggtcgc tctaatcact ccacggcagc 781 gcgctagagt gctggta (SEQ ID NO: 14516) .

[0295] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of CCCGGCGAGCATGAGG (SEQ ID NO: 14510). In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of SEQ ID NO: 14510. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTATCCCGGCGAGCATGAGG (SEQ ID NO: 14511). In certain embodiments, the piggyBac or piggyBac-like transposon comprises at least 16 contiguous nucleotides from SEQ ID NO: 14511. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of CCTCATGCTCGCCGGGTTAT (SEQ ID NO: 14512). In certain embodiments, the piggyBac or piggyBac-like transposon comprises at least 16 contiguous nucleotides from SEQ ID NO: 14512. In certain embodiments, the piggyBac or piggyBac- like transposon comprises one end comprising at least 16 contiguous nucleotides from SEQ ID NO: 14511 and one end comprising at least 16 contiguous nucleotides from SEQ ID NO: 14512. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14511 and SEQ ID NO: 14512. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCCGGCGAGCATGAGG (SEQ ID NO: 14513). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of CCTCATGCTCGCCGGGTTAA (SEQ ID NO: 14514).

[0296] In certain embodiments, the piggyBac or piggyBac-like transposon may have ends comprising SEQ ID NO: 14506 and SEQ ID NO: 14507, or a variant of either or both of these having at least 90% sequence identity to SEQ ID NO: 14506 or SEQ ID NO: 14507, and the piggyBac or piggyBac-like transposase has the sequence of SEQ ID NO: 14504 or SEQ ID NO: 14505, or a sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identity to SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a heterologous polynucleotide inserted between a pair of inverted repeats, where the transposon is capable of transposition by a piggyBac or piggyBac-like transposase having at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identity to SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the transposon comprises two transposon ends, each of which comprises SEQ ID NO: 14510 in inverted orientations in the two transposon ends. In certain embodiments, each inverted terminal repeat (ITR) is at least 90% identical to SEQ ID NO: 14510.

[0297] In certain embodiments, the piggyBac or piggyBac-like transposon is capable of insertion by a piggyBac or piggyBac-like transposase at the sequence 5'-TTAT-3 within a target nucleic acid. In certain embodiments, one end of the piggyBac or piggyBac-like transposon comprises at least 16 contiguous nucleotides from SEQ ID NO: 14506 and the other transposon end comprises at least 16 contiguous nucleotides from SEQ ID NO: 14507. In certain embodiments, one end of the piggyBac or piggyBac-like transposon comprises at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 contiguous nucleotides from SEQ ID NO: 14506 and the other transposon end comprises at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 contiguous nucleotides from SEQ ID NO: 14507.

[0298] In certain embodiments, the piggyBac or piggyBac-like transposon comprises transposon ends (each end comprising an ITR) corresponding to SEQ ID NO: 14506 and SEQ ID NO: 14507, and has a target sequence corresponding to 5'-TTAT3'. In certain embodiments, the piggyBac or piggyBac-like transposon also comprises a sequence encoding a transposase (e.g. SEQ ID NO: 14505). In certain embodiments, the piggyBac or piggyBac- like transposon comprises one transposon end corresponding to SEQ ID NO: 14506 and a second transposon end corresponding to SEQ ID NO: 14516. SEQ ID NO: 14516 is very similar to SEQ ID NO: 14507, but has a large insertion shortly before the ITR. Although the ITR sequences for the two transposon ends are identical (they are both identical to SEQ ID NO: 14510), they have different target sequences: the second transposon has a target sequence corresponding to 5'-TTAA-3', providing evidence that no change in ITR sequence is necessary to modify the target sequence specificity. The piggyBac or piggyBac-like transposase (SEQ ID NO: 14504), which is associated with the 5'-TTAA-3' target site differs from the 5'-TTAT-3'-associated transposase (SEQ ID NO: 14505) by only 4 amino acid changes (D322Y, S473C, A507T, H582R). In certain embodiments, the piggyBac or piggyBac-like transposase (SEQ ID NO: 14504), which is associated with the 5'-TTAA-3' target site is less active than the 5 '-TTAT-3 '-associated piggyBac or piggyBac-like transposase (SEQ ID NO: 14505) on the transposon with 5'-TTAT-3' ends. In certain embodiments, piggyBac or piggyBac-like transposons with 5'-TTAA-3' target sites can be converted to piggyBac or piggyBac-like transposases with 5'-TTAT-3 target sites by replacing 5'-TTAA-3' target sites with 5'-TTAT-3'. Such transposons can be used either with a piggyBac or piggyBac-like transposase such as SEQ ID NO: 14504 which recognizes the 5'-TTAT-3' target sequence, or with a variant of a transposase originally associated with the 5'-TTAA-3' transposon. In certain embodiments, the high similarity between the 5'-TTAA-3' and 5'-TTAT-3' piggyBac or piggyBac-like transposases demonstrates that very few changes to the amino acid sequence of a piggyBac or piggyBac-like transposase alter target sequence specificity. In certain embodiments, modification of any piggyBac or piggyBac-like transposon-transposase gene transfer system, in which 5'-TTAA-3' target sequences are replaced with 5'-TTAT-3'-target sequences, the ITRs remain the same, and the transposase is the original piggyBac or piggyBac-like transposase or a variant thereof resulting from using a low-level mutagenesis to introduce mutations into the transposase. In certain embodiments, piggyBac or piggyBac-like transposon transposase transfer systems can be formed by the modification of a 5'-TTAT-3'-active piggyBac or piggyBac-like transposon-transposase gene transfer systems in which 5'-TTAT-3' target sequences are replaced with 5'-TTAA-3'-target sequences, the ITRs remain the same, and the piggyBac or piggyBac-like transposase is the original transposase or a variant thereof.

[0299] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Bombyx mori. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt 61 ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga 121 ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac 181 ctgtttcggg tatgttatac cctgcctcat tgttgacgta t (SEQ ID NO: 14577).

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttaagaaaa agattaataa ataataataa tttcataatt aaaaacttct ttcattgaat 61 gccattaaat aaaccattat tttacaaaat aagatcaaca taattgagta aataataata 121 agaacaatat tatagtacaa caaaatatgg gtatgtcata ccctgccaca ttcttgatgt 181 aacttttttt cacctcatgc tcgccggg (SEQ ID NO: 14578) .

In certain embodiments, the transposon comprises at least 16 contiguous bases from SEQ ID NO: 14577 and at least 16 contiguous bases from SEQ ID NO: 14578, and inverted terminal repeats that are at least 87% identical to CCCGGCGAGC ATGAGG (SEQ ID NO: 14510). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt 61 ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga 121 ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac 181 ctgtttcggg tatgttatac cctgcctcat tgttgacgta ttttttttat gtaatttttc 241 cgattattaa tttcaactgt tttattggta tttttatgtt atccattgtt ctttttttat 301 gatttactgt atcggttgtc tttcgttcct ttagttgagt ttttttttat tattttcagt 361 ttttgatcaa a (SEQ ID NO: 14595) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct 61 ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt 121 gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa 181 taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataaac cattatttta 241 caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa 301 atatgggtat gtcataccct gccacattct tgatgtaact ttttttcacc tcatgctcgc 361 cggg (SEQ ID NO: 14596) .

[0300] In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14595 and SEQ ID NO: 14596, and is transposed by the piggyBac or piggyBac-like transposase of SEQ ID NO: 14505. In certain embodiments, the ITRs of SEQ ID NO: 14595 and SEQ ID: 14596 are not flanked by a 5 '-TTAA-3 ' sequence. In certain embodiments, the ITRs of SEQ ID NO: 14595 and SEQ ID: 14596 are flanked by a 5'-TTAT-3 ' sequence.

[0301] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt 61 ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga

121 ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac

181 ctgtttcggg tatgttatac cctgcctcat tgttgacgta ttttttttat gtaatttttc

241 cgattattaa tttcaactgt tttattggta tttttatgtt atccattgtt ctttttttat

301 g (SEQ ID NO: 14597) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 cagggtatct cataccctgg taaaatttta aagttgtgta ttttataaaa ttttcgtctg

61 acaacactag cgcgctcagt agctggaggc aggagcgtgc gggaggggat agtggcgtga

121 tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc aaacctgttt cgggtatgtt

181 ataccctgcc tcattgttga cgtatttttt ttatgtaatt tttccgatta ttaatttcaa

241 ctgttttatt ggtattttta tgttatccat tgttcttttt ttatg (SEQ ID NO: 14598) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 cagggtatct cataccctgg taaaatttta aagttgtgta ttttataaaa ttttcgtctg

61 acaacactag cgcgctcagt agctggaggc aggagcgtgc gggaggggat agtggcgtga

121 tcgcagtgtg gcacgggaca ccggcgagat attcgtgtgc aaacctgttt cgggtatgtt

181 ataccctgcc tcattgttga cgtat (SEQ ID NO: 14599) .

In certain embodiments, the left end of the piggyBac or piggyBac-like transposon comprises a sequence of SEQ ID NO: 14577, SEQ ID NO: 14595, or SEQ ID NOs: 14597-14599. In certain embodiments, the left end of the piggyBac or piggyBac-like transposon is preceded by a left target sequence.

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tcatattttt agtttaaaaa aataattata tgttttataa tgaaaagaat ctcattatct

61 ttcagtatta ggttgattta tattccaaag aataatattt ttgttaaatt gttgattttt

121 gtaaacctct aaatgtttgt tgctaaaatt actgtgttta agaaaaagat taataaataa

181 taataatttc ataattaaaa acttctttca ttgaatgcca ttaaataaac cattatttta

241 caaaataaga tcaacataat tgagtaaata ataataagaa caatattata gtacaacaaa

301 atatgggtat gtcataccct gccacattct tgatgtaact ttttttcacc tcatgctcgc

361 cggg (SEQ ID NO: 14600) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttaagaaaa agattaataa ataataataa tttcataatt aaaaacttct ttcattgaat

61 gccattaaat aaaccattat tttacaaaat aagatcaaca taattgagta aataataata

121 agaacaatat tatagtacaa caaaatatgg gtatgtcata ccctgccaca ttcttgatgt

181 aacttttttt ca (SEQ ID NO: 14601) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 cccggcgagc atgaggcagg gtatctcata ccctggtaaa attttaaagt tgtgtatttt

61 ataaaatttt cgtctgacaa cactagcgcg ctcagtagct ggaggcagga gcgtgcggga

121 ggggatagtg gcgtgatcgc agtgtggcac gggacaccgg cgagatattc gtgtgcaaac

181 ctgtttcggg tatgttatac cctgcctcat tgttgacgta ttttttttat gtaatttttc

241 cgattattaa tttcaactgt tttattggta tttttatgtt atccattgtt ctttttttat

301 gatttactgt atcggttgtc tttcgttcct ttagttgagt ttttttttat tattttcagt

361 ttttgatcaa a (SEQ ID NO: 14602) .

[0302] In certain embodiments, the right end of the piggyBac or piggyBac-like transposon comprises a sequence of SEQ ID NO: 14578, SEQ ID NO: 14596, or SEQ ID NOs: 14600- 14601. In certain embodiments, the right end of the piggyBac or piggyBac-like transposon is followed by a right target sequence. In certain embodiments, the transposon is transposed by the transposase of SEQ ID NO: 14505. In certain embodiments, the left and right ends of the piggyBac or piggy Bac-like transposon share a 16 bp repeat sequence of SEQ ID NO: 14510 in inverted orientation and immediately adjacent to the target sequence. In certain embodiments, the left transposon end begins with SEQ ID NO: 14510, and the right transposon end ends with the reverse complement of SEQ ID NO: 14510, 5'- CCTCATGCTCGCCGGG-3' (SEQ ID NO: 14603). In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR with at least 93%, at least 87%, or at least 81% or any percentage in between identity to SEQ ID NO: 14510 or SEQ ID NO: 14603. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a target sequence followed by a left transposon end comprising a sequence selected from SEQ ID NOs: 88, 105 or 107 and a right transposon end comprising SEQ ID NO: 14578 or 106 followed by a target sequence, in certain embodiments, the piggyBac or piggyBac like transposon comprises one end that comprises a sequence that is at least 90%, at least 95% or at least 99% or any percentage in between identical to SEQ ID NO: 14577 and one end that comprises a sequence that is at least 90%, at least 95% or at least 99% or any percentage in between identical to SEQ ID NO: 14578. In certain embodiments, one transposon end comprises at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14577 and one transposon end comprises at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14578.

[0303] In certain embodiments, the piggyBac or piggyBac-like transposon comprises two transposon ends wherein each transposon ends comprises a sequence that is at least 81% identical, at least 87% identical or at least 93% identical or any percentage in between identical to SEQ ID NO: 14510 in inverted orientation in the two transposon ends. One end may further comprise at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14599, and the other end may further comprise at least 14, at least 16, at least 18 or at least 20 contiguous bases from SEQ ID NO: 14601. The piggyBac or piggyBac-like transposon may be transposed by the transposase of SEQ ID NO: 14505, and the transposase may optionally be fused to a nuclear localization signal.

[0304] In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14595 and SEQ ID NO: 14596 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14597 and SEQ ID NO: 14596 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO:

14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14595 and SEQ ID NO: 14578 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO: 14505. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14602 and SEQ ID NO: 14600 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14504 or SEQ ID NO:

14505.

[0305] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left end comprising 1, 2, 3, 4, 5, 6, or 7 sequences selected from ATGAGGCAGGGTAT (SEQ ID NO: 14614), ATACCCTGCCTCAT (SEQ ID NO: 14615), GGCAGGGTAT (SEQ ID NO: 14616), ATACCCTGCC (SEQ ID NO: 14617), TAAAATTTTA (SEQ ID NO: 14618), ATTTTATAAAAT (SEQ ID NO: 14619), TCATACCCTG (SEQ ID NO: 14620) and TAAATAATAATAA (SEQ ID NO: 14621). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a right end comprising 1, 2 or 3 sequences selected from SEQ ID NO: 14617, SEQ ID NO: 14620 and SEQ ID NO: 14621.

[0306] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Xenopus tropicalis. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV 61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL 121 FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN 181 SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID 241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF 301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT 361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE 421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY 481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP 541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY (SEQ ID NO: 14517) .

[0307] In some embodiments, the piggyBac or piggyBac-like transposase is a hyperactive variant of SEQ ID NO: 14517. In certain embodiments, the piggyBac or piggyBac-like transposase is an integration defective variant of SEQ ID NO: 14517. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MAKRFYSAEE AAAHCMAPSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV 61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL 121 FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN 181 SLESYWNTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPDHD RLHKLRPLID 241 SLSERFAAVY TPCQNICIDE SLLLFKGRLR FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF 301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT 361 PACGTINRTR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE 421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT SAWYKKVGIY LIQMALRNSY 481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMLP SDNVARLIGK HFIDTLPPTP 541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY (SEQ ID NO: 14518) .

[0308] In certain embodiments, the piggyBac or piggyBac-like transposase is isolated or derived from Xenopus tropicalis. In certain embodiments, the piggyBac or piggyBac-like transposase is a hyperactive piggyBac or piggyBac-like transposase. In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence at least 90% identical to:

1 MAKRFYSAEE AAAHCSASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV 61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL 121 FMTEAILQDM VLYTNVYAEQ YLTQNPLTRG ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN 181 SIESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID 241 SLSERFANVY TPCQNICIDE SLMLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSTGYTSYF 301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT 361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE 421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY 481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPD SDNVARLIGK HFIDTLPPTP 541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY (SEQ ID NO: 14572) .

[0309] In certain embodiments, piggyBac or piggyBac-like transposase is a hyperactive piggyBac or piggyBac-like transposase. A hyperactive piggyBac or piggyBac-like transposase is a transposase that is more active than the naturally occurring variant from which it is derived. In certain embodiments, a hyperactive piggyBac or piggyBac-like transposase is more active than the transposase of SEQ ID NO: 14517. In certain

embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MAKRFYSAEE AAAHCSASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV 61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL 121 FMTEAILQDM VLYTNVYAEQ YLTQNPLTRG ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN 181 SIESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID 241 SLSERFANVY TPCQNICIDE SLMLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSTGYTSYF 301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT 361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE 421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY 481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPD SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY (SEQ ID NO:

14572).

[0310] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNPLTRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF 301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY (SEQ ID NO:

14624) .

[0311] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLKIPVFSAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY (SEQ ID NO:

14625) .

[0312] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MAKRFYSAEE AAAHCMASSS EQTSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNPLTRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SIESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241 SLSERFANVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRKPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY (SEQ ID NO: 14627) .

[0313] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MAKRFYSAEE AAAHCSASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNPLTRG ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241 SLSERFANVY TPCQNICIDE SLMLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSTGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY (SEQ ID NO: 14628) .

[0314] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNPLTRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLSIPVFGAT MSRNRYQLLL RFLHFNNNAT AVPPDQPGHD RLHKLRPLID

241 SLSERFANVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLNT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE 421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RHWYKKVGIY LIQMALRNSY 481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP 541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCRKPCF EIYHTQLHY (SEQ ID NO:

149) .

[0315] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution at a position selected from amino acid 6, 7, 16, 19, 20, 21, 22, 23, 24, 26, 28, 31, 34, 67, 73, 76, 77, 88, 91, 141, 145, 146, 148, 150, 157, 162, 179, 182, 189, 192, 193, 196, 198, 200, 210, 212, 218, 248, 263, 270, 294, 297, 308, 310, 333, 336, 354, 357, 358, 359, 377, 423, 426, 428, 438, 447, 450, 462, 469, 472, 498, 502, 517, 520, 523, 533, 534, 576, 577, 582, 583 or 587 (relative to SEQ ID NO: 14517). In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises an amino acid substitution of Y6C, S7G, M16S, S19G, S20Q, S20G, S20D, E21D, E22Q, F23T, F23P, S24Y, S26V, S28Q, V31K, A34E, L67A, G73H, A76V, D77N, P88A, N91D, Y141Q, Y141A, N145E, N145V, P146T, P146V, P146K, P148T, P148H, Y150G, Y150S, Y150C, H157Y, A162C, A179K, L182I, L182V, T189G, L192H, S 193N, S193K, V196I, S198G, T200W, L210H, F212N, N218E, A248N, L263M, Q270L, S294T, T297M, S308R, L310R, L333M, Q336M, A354H, C357V, L358F, D359N, L377I, V 423H, P426K, K428R, S438A, T447G, T447A, L450V, A462H, A462Q, I469V, I472L, Q498M, L502V, E5171, P520D, P520G, N523S, I533E, D534A, F576R, F576E, K577I, I582R, Y583F, L587Y or L587W, or any combination thereof including at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or all of these mutations (relative to SEQ ID NO: 14517).

[0316] In certain embodiments, the hyperactive piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of A2X, K3X, R4X, F5X, Y6X, S7X, A11X, A13X, C15X, M16X, A17X, S18X, S19X, S20X, E21X, E22X, F23X, S24X, G25X, 26X, D27X, S28X, E29X, E42X, E43X, S44X, C46X, S47X, S48X, S49X, T50X, V51X, S52X, A53X, L54X, E55X, E56X, P57X, M58X, E59X, E62X, D63X, V64X, D65X, D66X, L67X, E68X, D69X, Q70X, E71X, A72X, G73X, D74X, R75X, A76X, D77X, A78X, A79X, A80X, G81X, G82X, E83X, P84X, A85X, W86X, G87X, P88X, P89X, C90X, N91X, F92X, P93X, E95X, I96X, P97X, P98X, F99X, T100X, T101X, P103X, G104X, V105X, K106X, V107X, D108X, T109X, N111X, P114X, I115X, N116X, F117X, F118X, Q119X, M122X, T123X, E124X, A125X, I126X, L127X, Q128X, D129X, M130X, L132X, Y133X, V126X, Y127X, A138X, E139X, Q140X, Y141X, L142X, Q144X, N145X, P146X, L147X, P148X, Y150X, A151X, A155X, H157X, P158X , I161X, A162X, V168X, T171X, L172X, A173X, M174X, I177X, A179X, L182X, D187X, T188X, T189X, T190X, L192X, S193X, I194X, P195X, V196X, S198X, A199X, T200X, S202X, L208X, L209X, L210X, R211X, F212X, F215X, N217X, N218X, A219X, T220X, A221X, V222X, P224X, D225X, Q226X, P227X, H229X, R231X, H233X, L235X, P237X, I239X, D240X, L242X, S243X, E244X, R244X, F246X, A247X, A248X, V249X, Y250X, T251X, P252X, C253X, Q254X, I256X, C257X, I258X, D259X, E260X, S261X, L262X, L263X, L264X, F265X, K266X, G267X, R268X, L269X, Q270X, F271X, R272X, Q273X, Y274X, I275X, P276X, S277X, K278X, R279X, A280X, R281X, Y282X, G283X, I284X, K285X, F286X, Y287X, K288X, L289X, C290X, E291X, S292X, S293XS294X, G295X, Y296X, T297X, S298X, Y299X, F300X, E304X, L310X, P313X, G314X, P316X, P317X, D318X, L319X, T320X, V321X, K324X, E328X, I330X, S331X, P332X, L333X, L334X, G335X, Q336X, F338X, L340X, D343X, N344X, F345X, Y346X, S347X, L351X, F352X, A354X, L355X, Y356X, C357X, L358X, D359X, T360X, R422X, Y423X, G424X, P426X, K428X, N429X, K430X, P431X, L432X, S434X, K435X, E436X, S438X, K439X, Y440X, G443X, R446X, T447X, L450X, Q451X, N455X, T460X, R461X, A462X, K465X, V467X, G468X, I469X, Y470X, L471X, I472X, M474X, A475X, L476X, R477X, S479X, Y480X, V482XY483X, K484X, A485X, A486X, V487X, P488X, P490X, K491X, S493X, Y494X, Y495X, K496X, Y497T, Q498X, L499X, Q500X, I501X, L502X, P503X, A504X, L505X, L506X, F507X, G508X, G509X, V510X, E511X, E512X, Q513X, T514X, V515X, E517X, M518X, P519X, P520X, S521X, D522X, N523X, V524X, A525X, L527X, I528X, K530X, H531X, F532X, I533X, D534X, T535X, L536X, T539X, P540X,Q546X, K550X, R553X, K554X, R555X, G556X, I557X, R558X, R559X, D560X, T561X, Y564X, P566X, K567X, P569X, R570X, N571X, L574X, C575X, F576X, K577X, P578X, F580X, E581X, I582X, Y583X, T585X, Q586X, L587X, H588X or Y589X (relative to SEQ ID NO: 14517). A list of hyperactive amino acid substitutions can be found in US patent No. 10,041,077, the contents of which are incorporated by reference in their entirety.

[0317] In certain embodiments, the piggyBac or piggyBac-like transposase is integration deficient. In certain embodiments, an integration deficient piggyBac or piggyBac-like transposase is a transposase that can excise its corresponding transposon, but that integrates the excised transposon at a lower frequency than a corresponding naturally occurring transposase. In certain embodiments, the piggyBac or piggyBac-like transposase is an integration deficient variant of SEQ ID NO: 14517. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase is deficient relative to SEQ ID NO: 14517.

[0318] In certain embodiments, the piggyBac or piggyBac-like transposase is active for excision but deficient in integration. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of :

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRVDAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLSIPVFSAT MSRNRYQLLL KFLHFNNEAT AVPPDQPGHD RLHKLRPLID

241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR (SEQ ID NO: 14605) .

[0319] In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQVPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLNIPVFSAT MSRNRYQLLL RFLEFNNEAT AVPPDQPGHD RLHKLRPLID

241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHY (SEQ ID NO: 14604) .

[0320] In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQNVLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNDAT AVPPDQPGHD RLHKLRPLID

241 SLTERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP

541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR (SEQ ID NO: 14611) .

[0321] In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14611. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV

61 DEDVDDLEDQ EAGDRADAAP GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL

121 FMTEAILQDM VLYTNVYAEQ YLTQVPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN

181 SLESYWDTTT VLSIPVFSAT MSRNRYQLLL RFLHFNNEAT AVPPDQPGHD RLHKLRPLID

241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF

301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT

361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE

421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY

481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP 541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR (SEQ ID NO: 14612) .

[0322] In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14612. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises a sequence that is at least 90% identical to a sequence of:

1 MAKRFYSAEE AAAHCMASSS EEFSGSDSEY VPPASESDSS TEESWCSSST VSALEEPMEV 61 DEDVDDLEDQ EAGDRADAAA GGEPAWGPPC NFPPEIPPFT TVPGVKVDTS NFEPINFFQL 121 FMTEAILQDM VLYTNVYAEQ YLTQVPLPRY ARAHAWHPTD IAEMKRFVGL TLAMGLIKAN 181 SLESYWDTTT VLNIPVFSAT MSRNRYQLLL RFLEFNNNAT AVPPDQPGHD RLHKLRPLID 241 SLSERFAAVY TPCQNICIDE SLLLFKGRLQ FRQYIPSKRA RYGIKFYKLC ESSSGYTSYF 301 LIYEGKDSKL DPPGCPPDLT VSGKIVWELI SPLLGQGFHL YVDNFYSSIP LFTALYCLDT 361 PACGTINRNR KGLPRALLDK KLNRGETYAL RKNELLAIKF FDKKNVFMLT SIHDESVIRE 421 QRVGRPPKNK PLCSKEYSKY MGGVDRTDQL QHYYNATRKT RAWYKKVGIY LIQMALRNSY 481 IVYKAAVPGP KLSYYKYQLQ ILPALLFGGV EEQTVPEMPP SDNVARLIGK HFIDTLPPTP 541 GKQRPQKGCK VCRKRGIRRD TRYYCPKCPR NPGLCFKPCF EIYHTQLHYG RR (SEQ ID NO: 14613) .

[0323] In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14613. In certain embodiments, the integration deficient piggyBac or piggyBac-like transposase comprises an amino acid substitution wherein the Asn at position 218 is replaced by a Glu or an Asp (N218D or N218E) (relative to SEQ ID NO: 14517).

[0324] In certain embodiments, the excision competent, integration deficient piggyBac or piggyBac-like transposase comprises one or more substitutions of an amino acid that is not wild type, wherein the one or more substitutions a for wild type amino acid comprises a substitution of A2X, K3X, R4X, F5X, Y6X, S7X, A8X, E9X, E10X, A11X, A12X, A13X, H14X, C15X, M16X, A17X, S18X, S 19X, S20X, E21X, E22X, F23X, S24X, G25X, 26X, D27X, S28X, E29X, V31X, P32X, P33X, A34X, S35X, E36X, S37X, D38X, S39X, S40X, T41X, E42X, E43X, S44X, W45X, C46X, S47X, S48X, S49X, T50X, V51X, S52X, A53X, L54X, E55X, E56X, P57X, M58X, E59X, V60X, M122X, T123X, E124X, A125X, L127X, Q128X, D129X, L132X, Y133X, V126X, Y127X, E139X, Q140X, Y141X, L142X, T143X, Q144X, N145X, P146X, L147X, P148X, R149X, Y150X, A151X, H154X, H157X, P158X, T159X, D160X, I161X, A162X, E163X, M164X, K165X, R166X, F167X, V168X, G169X, L170X, T171X, L172X, A173X, M174X, G175X, L176X, I177X, K178X, A179X, N180X, S181X, L182X, S184X, Y185X, D187X, T188X, T189X, T190X, V191X, L192X, S193X, I194X, P195X, V196X, F197X, S198X, A199X, T200X, M201X, S202X, R203X, N204X, R205X, Y206X, Q207X, L208X, L209X, L210X, R211X, F212X, L213X, H241X, F215X, N216X, N217X, N218X, A219X, T220X, A221X, V222X, P223X, P224X, D225X, Q226X, P227X, G228X, H229X, D230X, R231X, H233X, K234X, L235X, R236X, L238X, I239X, D240X, L242X, S243X, E244X, R244X, F246X, A247X, A248X, V249X, Y250X, T251X, P252X, C253X, Q254X, N255X, I256X, C257X, I258X, D259X, E260X, S261X, L262X, L263X, L264X, F265X, K266X, G267X, R268X, L269X, Q270X, F271X, R272X, Q273X, Y274X, I275X, P276X, S277X, K278X, R279X, A280X, R281X, Y282X, G283X, I284X, K285X, F286X, Y287X, K288X, L289X, C290X, E291X, S292X, S293X, S294X, G295X, Y296X, T297X, S298X, Y299X, F300X, I302X, E304X, G305X,K306X, D307X, S308X, K309X, L310X, D311X, P312X, P313X, G314X, C315X, P316X, P317X, D318X, L319X, T320X, V321X, S322X, G323X, K324X, I325X, V326X, W327X, E328X, L329X, I330X, S331X, P332X, L333X, L334X, G335X, Q336X, F338X, H339X, L340X, V342X, N344X, F345X, Y346X, S347X, S348X, I349X, L351X, T353X, A354X, Y356X, C357X, L358X, D359X, T360X, P361X, A362X, C363X, G364X, I366X, N367X, R368X, D369X, K371X, G372X, L373X, R375X, A376X, L377X, L378X, D379X, K380X, K381X, L382X, N383X, R384XG385X, T387X, Y388X, A389X, L390X, K392X, N393X, E394X, A397X, K399X, F400X, F401X, D402X, N405X, L406X, L409X, R422X, Y423X, G424X, E425X, P426X, K428X, N429X, K430X, P431X, L432X, S434X, K435X, E436X, S438X, K439X, Y440X, G442X, G443X, V444X, R446X, T447X, L450X, Q451X, H452X, N455X, T457X, R458X, T460X, R461X, A462X, Y464X, K465X, V467X, G468X, I469X, L471X, I472X, Q473X, M474X, L476X, R477X, N478X, S479X, Y480X, V482XY483X, K484X, A485X, A486X, V487X, P488X, G489X, P490X, K491X, L492X, S493X, Y494X, Y495X, K496X, Q498X, L499X, Q500X, I501X, L502X, P503X, A504X, L505X, L506X, F507X, G508X, G509X, V510X, E511X, E512X, Q513X, T514X, V515X, E517X, M518X, P519X, P520X, S521X, D522X, N523X, V524X, A525X, L527X, I528X, G529X, K530X, F532X, I533X, D534X, T535X, L536X, P537X, P538X, T539X, P540X, G541X, F542X, Q543X, R544X, P545X, Q546X, K547X, G548X, C549X, K550X, V551X, C552X, R553X, K554X, R555X, G556X, I557X, R558X, R559X, D560X, T561X, R562X, Y563X, Y564X, C565X, P566X, K567X, C568X, P569X, R570X, N571X, P572X, G573X, L574X, C575X, F576X, K577X, P578X, C579X, F580X, E581X, I582X, Y583X, H584X, T585X, Q586X, L587X, H588X or Y589X (relative to SEQ ID NO: 14517). A list of excision competent, integration deficient amino acid substitutions can be found in US patent No. 10,041,077, the contents of which are incorporated by reference in their entirety.

[0325] In certain embodiments, the piggyBac or piggyBac-like transposase is fused to a nuclear localization signal. In certain embodiments, SEQ ID NO: 14517 or SEQ ID NO: 14518 is fused to a nuclear localization signal. In certain embodiments, the amino acid sequence of the piggyBac or piggyBac like transposase fused to a nuclear localization is encoded by a polynucleotide sequence comprising:

1 atggcaccca aaaagaaacg taaagtgatg gccaaaagat tttacagcgc cgaagaagca

6 1 gcagcacatt gcatggcatc gtcatccgaa gaattctcgg ggagcgattc cgaatatgtc 12 1 ccaccggcct cggaaagcga ttcgagcact gaggagtcgt ggtgttcctc ctcaactgtc 18 1 tcggctcttg aggagccgat ggaagtggat gaggatgtgg acgacttgga ggaccaggaa 24 1 gccggagaca gggccgacgc tgccgcggga ggggagccgg cgtggggacc tccatgcaat 30 1 tttcctcccg aaatcccacc gttcactact gtgccgggag tgaaggtcga cacgtccaac 36 1 ttcgaaccga tcaatttctt tcaactcttc atgactgaag cgatcctgca agatatggtg 42 1 ctctacacta atgtgtacgc cgagcagtac ctgactcaaa acccgctgcc tcgctacgcg 48 1 agagcgcatg cgtggcaccc gaccgatatc gcggagatga agcggttcgt gggactgacc 54 1 ctcgcaatgg gcctgatcaa ggccaacagc ctcgagtcat actgggatac cacgactgtg 60 1 cttagcattc cggtgttctc cgctaccatg tcccgtaacc gctaccaact cctgctgcgg 66 1 ttcctccact tcaacaacaa tgcgaccgct gtgccacctg accagccagg acacgacaga 72 1 ctccacaagc tgcggccatt gatcgactcg ctgagcgagc gattcgccgc ggtgtacacc 78 1 ccttgccaaa acatttgcat cgacgagtcg cttctgctgt ttaaaggccg gcttcagttc 84 1 cgccagtaca tcccatcgaa gcgcgctcgc tatggtatca aattctacaa actctgcgag 90 1 tcgtccagcg gctacacgtc atacttcttg atctacgagg ggaaggactc taagctggac 96 1 ccaccggggt gtccaccgga tcttactgtc tccggaaaaa tcgtgtggga actcatctca 102 1 cctctcctcg gacaaggctt tcatctctac gtcgacaatt tctactcatc gatccctctg 108 1 ttcaccgccc tctactgcct ggatactcca gcctgtggga ccattaacag aaaccggaag 114 1 ggtctgccga gagcactgct ggataagaag ttgaacaggg gagagactta cgcgctgaga 120 1 aagaacgaac tcctcgccat caaattcttc gacaagaaaa atgtgtttat gctcacctcc 126 1 atccacgacg aatccgtcat ccgggagcag cgcgtgggca ggccgccgaa aaacaagccg 132 1 ctgtgctcta aggaatactc caagtacatg gggggtgtcg accggaccga tcagctgcag 138 1 cattactaca acgccactag aaagacccgg gcctggtaca agaaagtcgg catctacctg 144 1 atccaaatgg cactgaggaa ttcgtatatt gtctacaagg ctgccgttcc gggcccgaaa 150 1 ctgtcatact acaagtacca gcttcaaatc ctgccggcgc tgctgttcgg tggagtggaa 156 1 gaacagactg tgcccgagat gccgccatcc gacaacgtgg cccggttgat cggaaagcac 162 1 ttcattgata ccctgcctcc gacgcctgga aagcagcggc cacagaaggg atgcaaagtt 168 1 tgccgcaagc gcggaatacg gcgcgatacc cgctactatt gcccgaagtg cccccgcaat 174 1 cccggactgt gtttcaagcc ctgttttgaa atctaccaca cccagttgca ttac (SEQ 14626) .

[0326] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Xenopus tropicalis. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaacctttt tactgccaat gacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg 61 ccaacgacgc gtcccatacg ttgttggcat tttaagtctt ctctctgcag cggcagcatg 121 tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg 181 ctgtc (SEQ ID NO: 14519) .

[0327] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa 61 ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg 121 taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa 181 actgtctggc aatacaagtt ccactttgac caaaacggct ggcagtaaaa gggttaa (SEQ ID NO: 14520) .

[0328] In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14519 and SEQ ID NO: 14520. In certain embodiments, the piggyBac or piggyBac- like transposon comprises a sequence of: 1 ttaacccttt gcctgccaat cacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg

61 ccaacgacgc gtcccatacg ttgttggcat tttaagtctt ctctctgcag cggcagcatg

121 tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg 181 ctgtc (SEQ ID NO: 14521) .

[0329] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61 ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg 121 taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa 181 actgtctggc aatacaagtt ccactttggg acaaatcggc tggcagtgaa agggttaa (SEQ ID NO: 14522) .

[0330] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaacctttt tactgccaat gacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg 61 ccaacgacgc gtcccatacg ttgttggcat tttaattctt ctctctgcag cggcagcatg 121 tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg 181 ctgtc (SEQ ID NO: 14523) .

[0331] In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14520 and SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14522 and SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides from SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end comprising at least 14, 16, 18, 20, 30 or 40 contiguous nucleotides from SEQ ID NO:

14520 or SEQ ID NO: 14522. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14519, SEQ ID NO:

14521 or SEQ ID NO: 14523. In certain embodiments, the piggyBac or piggyBac-like transposon comprises one end with at least 90% identity to SEQ ID NO: 14520 or SEQ ID NO: 14522. In one embodiment, one transposon end is at least 90% identical to SEQ ID NO: 14519 and the other transposon end is at least 90% identical to SEQ ID NO: 14520.

[0332] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCTTTTTACTGCCA (SEQ ID NO: 14524). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCCTTTGCCTGCCA (SEQ ID NO: 14526). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCYTTTTACTGCCA (SEQ ID NO: 14527). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of

TGGCAGTAAAAGGGTTAA (SEQ ID NO: 14529). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TGGC AGTGAAAGGGTTAA (SEQ ID NO: 14531). In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of TTAACCYTTTKMCTGCCA (SEQ ID NO: 14533). In certain embodiments, one end of the piggyBac or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14524, SEQ ID NO: 14526 and SEQ ID NO: 14527. In certain embodiments, one end of the piggyBac™ (PB) or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14529 and SEQ ID NO: 14531. In certain embodiments, each inverted terminal repeat of the piggyBac or piggyBac-like transposon comprises a sequence of ITR sequence of CCYTTTKMCTGCCA (SEQ ID NO: 14563). In certain embodiments, each end of the piggyBac™ (PB) or piggyBac-like transposon comprises SEQ ID NO: 14563 in inverted orientations. In certain embodiments, one ITR of the piggyBac or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14524, SEQ ID NO: 14526 and SEQ ID NO: 14527. In certain embodiments, one ITR of the piggyBac or piggyBac-like transposon comprises a sequence selected from SEQ ID NO: 14529 and SEQ ID NO: 14531. In certain embodiments, the piggyBac or piggyBac like transposon comprises SEQ ID NO: 14533 in inverted orientation in the two transposon ends.

[0333] In certain embodiments, The piggyBac or piggyBac-like transposon may have ends comprising SEQ ID NO: 14519 and SEQ ID NO: 14520 or a variant of either or both of these having at least 90% sequence identity to SEQ ID NO: 14519 or SEQ ID NO: 14520, and the piggyBac or piggyBac-like transposase has the sequence of SEQ ID NO: 14517 or a variant showing at least %, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between sequence identity to SEQ ID NO: 14517 or SEQ ID NO: 14518. In certain embodiments, one piggyBac or piggyBac-like transposon end comprises at least 14 contiguous nucleotides from SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523, and the other transposon end comprises at least 14 contiguous nucleotides from SEQ ID NO: 14520 or SEQ ID NO: 14522. In certain embodiments, one transposon end comprises at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 22, at least 25, at least 30 contiguous nucleotides from SEQ ID NO: 14519, SEQ ID NO: 14521 or SEQ ID NO: 14523, and the other transposon end comprises at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 22, at least 25 or at least 30 contiguous nucleotides from SEQ ID NO: 14520 or SEQ ID NO:

14522.

[0334] In certain embodiments, the piggyBac or piggyBac-like transposase recognizes a transposon end with a left sequence corresponding to SEQ ID NO: 14519, and a right sequence corresponding to SEQ ID NO: 14520. It will excise the transposon from one DNA molecule by cutting the DNA at the 5'-TTAA-3' sequence at the left end of one transposon end to the 5'-TTAA-3' at the right end of the second transposon end, including any heterologous DNA that is placed between them, and insert the excised sequence into a second DNA molecule. In certain embodiments, truncated and modified versions of the left and right transposon ends will also function as part of a transposon that can be transposed by the piggyBac or piggyBac-like transposase. For example, the left transposon end can be replaced by a sequence corresponding to SEQ ID NO: 14521 or SEQ ID NO: 14523, the right transposon end can be replaced by a shorter sequence corresponding to SEQ ID NO: 14522. In certain embodiments, the left and right transposon ends share an 18 bp almost perfectly repeated sequence at their ends (5'-TTAACCYTTTKMCTGCCA: SEQ ID NO: 14533) that includes the 5'-TTAA-3' insertion site, which sequence is inverted in the orientation in the two ends. That is in SEQ ID NO: 14519 and SEQ ID NO: 14523 the left transposon end begins with the sequence 5'-TTAACCTTTTTACTGCCA-3' (SEQ ID NO: 14524), or in SEQ ID NO: 14521 the left transposon end begins with the sequence 5'- TTAACCCTTTGCCTGCCA-3' (SEQ ID NO: 14526); the right transposon ends with approximately the reverse complement of this sequence: in SEQ ID NO: 14520 it ends 5' TGGC AGTAAAAGGGTTAA-3 ' (SEQ ID NO: 14529), in SEQ ID NO: 14522 it ends 5'- TGGC AGTGAAAGGGTTAA-3 ' (SEQ ID NO: 14531.) One embodiment of the invention is a transposon that comprises a heterologous polynucleotide inserted between two transposon ends each comprising SEQ ID NO: 14533 in inverted orientations in the two transposon ends. In certain embodiments, one transposon end comprises a sequence selected from SEQ ID NOS: 14524, SEQ ID NO: 14526 and SEQ ID NO: 14527. In some embodiments, one transposon end comprises a sequence selected from SEQ ID NO: 14529 and SEQ ID NO: 14531.

[0335] In certain embodiments, the piggyBac™ (PB) or piggyBac-like transposon is isolated or derived from Xenopus tropicalis. In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 ccctttgcct gccaatcacg catgggatac gtcgtggcag taaaagggct taaatgccaa 61 cgacgcgtcc catacgtt (SEQ ID NO: 14573) .

[0336] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 cctgggtaaa ctaaaagtcc cctcgaggaa aggcccctaa agtgaaacag tgcaaaacgt 61 tcaaaaactg tctggcaata caagttccac tttgggacaa atcggctggc agtgaaaggg

(SEQ ID NO: 14574). [0337] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at least 16 contiguous bases from SEQ ID NO: 14573 or SEQ ID NO: 14574, and inverted terminal repeat of CCYTTTBMCTGCCA (SEQ ID NO: 14575).

[0338] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 ccctttgcct gccaatcacg catgggatac gtcgtggcag taaaagggct taaatgccaa 61 cgacgcgtcc catacgttgt tggcatttta agtcttctct ctgcagcggc agcatgtgcc 121 gccgctgcag agagtttcta gcgatgacag cccctctggg caacgagccg ggggggctgt 181 c (SEQ ID NO: 14579) .

[0339] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa 61 cgacgcgtcc catacgttgt tggcatttta attcttctct ctgcagcggc agcatgtgcc 121 gccgctgcag agagtttcta gcgatgacag cccctctggg caacgagccg ggggggctgt 181 c (SEQ ID NO: 14580) .

[0340] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa

61 cgacgcgtcc catacgttgt tggcatttta agtcttctct ctgcagcggc agcatgtgcc

121 gccgctgcag agagtttcta gcgatgacag cccctctggg caacgagccg ggggggctgt

181 c (SEQ ID NO: 14581) .

[0341] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa 61 cgacgcgtcc catacgttgt tggcatttta agtcttctct ctgcagcggc agcatgtgcc 121 gccgctgcag agag (SEQ ID NO: 14582) .

[0342] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 cctttttact gccaatgacg catgggatac gtcgtggcag taaaagggct taaatgccaa 61 cgacgcgtcc catacgttgt tggcatttta agtctt (SEQ ID NO: 14583).

[0343] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 ccctttgcct gccaatcacg catgggatac gtcgtggcag taaaagggct taaatgccaa 61 cgacgcgtcc catacgttgt tggcatttta agtctt (SEQ ID NO: 14584).

[0344] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 ttatcctttt tactgccaat gacgcatggg atacgtcgtg gcagtaaaag ggcttaaatg 61 ccaacgacgc gtcccatacg ttgttggcat tttaagtctt ctctctgcag cggcagcatg 121 tgccgccgct gcagagagtt tctagcgatg acagcccctc tgggcaacga gccggggggg 181 ctgtc (SEQ ID NO: 14585) . [0345] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61 ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121 taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181 actgtctggc aatacaagtt ccactttggg acaaatcggc tggcagtgaa aggg (SEQ ID NO: 14586) .

[0346] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left transposon end sequence selected from SEQ ID NO: 14573 and SEQ ID NOs: 14579-14585. In certain embodiments, the left transposon end sequence is preceded by a left target sequence. In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa 61 ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg 121 taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa 181 actgtctggc aatacaagtt ccactttgac caaaacggct ggcagtaaaa ggg (SEQ ID

NO: 14587) .

[0347] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 ttgttctgaa aaaaacaata tattgttttc ctgggtaaac taaaagtccc ctcgaggaaa 61 ggcccctaaa gtgaaacagt gcaaaacgtt caaaaactgt ctggcaatac aagttccact 121 ttgaccaaaa cggctggcag taaaaggg (SEQ ID NO: 14588) .

[0348] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 tttgcatttt tagacattta gaagcctata tcttgttaca gaattggaat tacacaaaaa

61 ttctaccata ttttgaaagc ttaggttgtt ctgaaaaaaa caatatattg ttttcctggg

121 taaactaaaa gtcccctcga ggaaaggccc ctaaagtgaa acagtgcaaa acgttcaaaa

181 actgtctggc aatacaagtt ccactttgac caaaacggct ggcagtaaaa gggttat (SEQ ID NO: 14589) .

[0349] In certain embodiments, the piggyBac or piggyBac-like transposon comprises at a sequence of:

1 ttgttctgaa aaaaacaata tattgttttc ctgggtaaac taaaagtccc ctcgaggaaa 61 ggcccctaaa gtgaaacagt gcaaaacgtt caaaaactgt ctggcaatac aagttccact 121 ttgggacaaa tcggctggca gtgaaaggg (SEQ ID NO: 14590).

[0350] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a right transposon end sequence selected from SEQ ID NO: 14574 and SEQ ID NOs: 14587-14590.

In certain embodiments, the right transposon end sequence is followed by a right target sequence. In certain embodiments, the left and right transposon ends share a 14 repeated sequence inverted in orientation in the two ends (SEQ ID NO: 14575) adjacent to the target sequence. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a left transposon end comprising a target sequence and a sequence that is selected from SEQ ID NOs: 14582-14584 and 14573, and a right transposon end comprising a sequence selected from SEQ ID NOs: 14588-14590 and 14574 followed by a right target sequence.

[0351] In certain embodiments, the left transposon end of the piggy Bac or piggy Bac-like transposon comprises

1 atcacgcatg ggatacgtcg tggcagtaaa agggcttaaa tgccaacgac gcgtcccata 61 cgtt

(SEQ ID NO: 14591), and an ITR. In certain embodiments, the left transposon end comprises

1 atgacgcatg ggatacgtcg tggcagtaaa agggcttaaa tgccaacgac gcgtcccata 61 cgttgttggc attttaagtc tt

(SEQ ID NO: 14592) and an ITR. In certain embodiments, the right transposon end of the piggyBac or piggyBac-like transposon comprises

1 cctgggtaaa ctaaaagtcc cctcgaggaa aggcccctaa agtgaaacag tgcaaaacgt 61 tcaaaaactg tctggcaata caagttccac tttgggacaa atcggc

(SEQ ID NO: 14593) and an ITR. In certain embodiments, the right transposon end comprises

1 ttgttctgaa aaaaacaata tattgttttc ctgggtaaac taaaagtccc ctcgaggaaa 61 ggcccctaaa gtgaaacagt gcaaaacgtt caaaaactgt ctggcaatac aagttccact 121 ttgaccaaaa cggc

(SEQ ID NO: 14594) and an ITR.

[0352] In certain embodiments, one transposon end comprises a sequence that is at least 90%, at least 95%, at least 99% or any percentage in between identical to SEQ ID NO: 14573 and the other transposon end comprises a sequence that is at least 90%, at least 95%, at least 99% or any percentage in between identical to SEQ ID NO: 14574. In certain embodiments, one transposon end comprises at least 14, at least 16, at least 18, at least 20 or at least 25 contiguous nucleotides from SEQ ID NO: 14573 and one transposon end comprises at least 14, at least 16, at least 18, at least 20 or at least 25 contiguous nucleotides from SEQ ID NO: 14574. In certain embodiments, one transposon end comprises at least 14, at least 16, at least 18, at least 20 from SEQ ID NO: 14591, and the other end comprises at least 14, at least 16, at least 18, at least 20 from SEQ ID NO: 14593. In certain embodiments, each transposon end comprises SEQ ID NO: 14575 in inverted orientations.

[0353] In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence selected from of SEQ ID NO: 14573, SEQ ID NO: 14579, SEQ ID NO: 14581, SEQ ID NO: 14582, SEQ ID NO: 14583, and SEQ ID NO: 14588, and a sequence selected from SEQ ID NO: 14587, SEQ ID NO: 14588, SEQ ID NO: 14589 and SEQ ID NO: 14586 and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14517 or SEQ ID NO: 14518.

[0354] In certain embodiments, the piggyBac or piggyBac-like transposon comprises ITRs of CCCTTTGCCTGCCA (SEQ ID NO: 14622) (left ITR) and TGGCAGTGAAAGGG (SEQ ID NO: 14623) (right ITR) adjacent to the target sequences.

[0355] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Helicoverpa armigera. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MASRQRLNHD EIATILENDD DYSPLDSESE KEDCWEDDV WSDNEDAIVD FVEDTSAQED

61 PDNNIASRES PNLEVTSLTS HRIITLPQRS IRGKNNHVWS TTKGRTTGRT SAINIIRTNR

121 GPTRMCRNIV DPLLCFQLFI TDEI IHEIVK WTNVEIIVKR QNLKDISASY RDTNTMEIWA

181 LVGILTLTAV MKDNHLSTDE LFDATFSGTR YVSVMSRERF EFLIRCIRMD DKTLRPTLRS

241 DDAFLPVRKI WEIFINQCRQ NHVPGSNLTV DEQLLGFRGR CPFRMYIPNK PDKYGIKFPM

301 MCAAATKYMI DAIPYLGKST KTNGLPLGEF YVKDLTKTVH GTNRNITCDN WFTSIPLAKN

361 MLQAPYNLTI VGTIRSNKRE MPEEIKNSRS RPVGSSMFCF DGPLTLVSYK PKPSKMVFLL

421 SSCDENAVIN ESNGKPDMIL FYNQTKGGVD SFDQMCKSMS ANRKTNRWPM AVFYGMLNMA

481 FV SYIIYCH NKINKQEKPI SRKEFMKKLS IQLTTPWMQE RLQAPTLKRT LRDNITNVLK

541 NWPASSENI SNEPEPKKRR YCGVCSYKKR RMTKAQCCKC KKAICGEHNI DVCQDCI (SEQ ID NO: 14525) .

[0356] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Helicoverpa armigera. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaaccctag aagcccaatc tacgtaaatt tgacgtatac cgcggcgaaa tatctctgtc 61 tctttcatgt ttaccgtcgg atcgccgcta acttctgaac caactcagta gccattggga 121 cctcgcagga cacagttgcg tcatctcggt aagtgccgcc attttgttgt actctctatt 181 acaacacacg tcacgtcacg tcgttgcacg tcattttgac gtataattgg gctttgtgta 241 acttttgaat ttgtttcaaa ttttttatgt ttgtgattta tttgagttaa tcgtattgtt 301 tcgttacatt tttcatataa taataatatt ttcaggttga gtacaaa (SEQ ID NO:

14570 ) . In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 agactgtttt tttctaagag acttctaaaa tattattacg agttgattta attttatgaa 61 aacatttaaa actagttgat tttttttata attacataat tttaagaaaa agtgttagag 121 gcttgatttt tttgttgatt ttttctaaga tttgattaaa gtgccataat agtattaata 181 aagagtattt tttaacttaa aatgtatttt atttattaat taaaacttca attatgataa 241 ctcatgcaaa aatatagttc attaacagaa aaaaatagga aaactttgaa gttttgtttt 301 tacacgtcat ttttacgtat gattgggctt tatagctagt taaatatgat tgggcttcta 361 gggttaa (SEQ ID NO: 14528) .

[0357] in certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Pectinophora gossypiella. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MDLRKQDEKI RQWLEQDIEE DSKGESDNSS SETEDIVEME VHKNTSSESE VSSESDYEPV

61 CPSKRQRTQI IESEESDNSE SIRPSRRQTS RVIDSDETDE DVMSSTPQNI PRNPNVIQPS

121 SRFLYGKNKH KWSSAAKPSS VRTSRRNIIH FIPGPKERAR EVSEPIDIFS LFISEDMLQQ

181 WTFTNAEML IRKNKYKTET FTVSPTNLEE IRALLGLLFN AAAMKSNHLP TRMLFNTHRS

241 GTIFKACMSA ERLNFLIKCL RFDDKLTRNV RQRDDRFAPI RDLWQALISN FQKWYTPGSY

301 ITVDEQLVGF RGRCSFRMYI PNKPNKYGIK LVMAADVNSK YIVNAIPYLG KGTDPQNQPL

361 ATFFIKEITS TLHGTNRNIT MDNWFTSVPL ANELLMAPYN LTLVGTLRSN KREIPEKLKN

421 SKSRAIGTSM FCYDGDKTLV SYKAKSNKW FILSTIHDQP DINQETGKPE MIHFYNSTKG

481 AVDTVDQMCS SISTNRKTQR WPLCVFYNML NLSIINAYW YVYNNVRNNK KPMSRRDFVI

541 KLGDQLMEPW LRQRLQTVTL RRDIKVMIQD ILGESSDLEA PVPSVSNVRK IYYLCPSKAR

601 RMTKHRCIKC KQAICGPHNI DICSRCIE (SEQ ID NO: 14530) .

[0358] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Pectinophora gossypiella. In certain embodiments, the piggyBac or piggyBac- like transposon comprises a sequence of:

1 ttaaccctag ataactaaac attcgtccgc tcgacgacgc gctatgccgc gaaattgaag

61 tttacctatt attccgcgtc ccccgccccc gccgcttttt ctagcttcct gatttgcaaa

121 atagtgcatc gcgtgacacg ctcgaggtca cacgacaatt aggtcgaaag ttacaggaat

181 ttcgtcgtcc gctcgacgaa agtttagtaa ttacgtaagt ttggcaaagg taagtgaatg

241 aagtattttt ttataattat tttttaattc tttatagtga taacgtaagg tttatttaaa

301 tttattactt ttatagttat ttagccaatt gttataaatt ccttgttatt gctgaaaaat

361 ttgcctgttt tagtcaaaat ttattaactt ttcgatcgtt ttttag (SEQ ID NO:

14532 ) . In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttcactaag taattttgtt cctatttagt agataagtaa cacataatta ttgtgatatt

61 caaaacttaa gaggtttaat aaataataat aaaaaaaaaa tggtttttat ttcgtagtct

121 gctcgacgaa tgtttagtta ttacgtaacc gtgaatatag tttagtagtc tagggttaa (SEQ ID NO: 14571) .

[0359] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Ctenoplusia agnata. The piggyBac or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MASRQHLYQD EIAAILENED DYSPHDTDSE MEDCVTQDDV RSDVEDEMVD NIGNGTSPAS 61 RHEDPETPDP SSEASNLEVT LSSHRIIILP QRSIREKNNH IWSTTKGQSS GRTAAINIVR 121 TNRGPTRMCR NIVDPLLCFQ LFIKEEIVEE IVKWTNVEMV QKRVNLKDIS ASYRDTNEME 181 IWAIISMLTL SAVMKDNHLS TDELFNVSYG TRYVSVMSRE RFEFLLRLLR MGDKLLRPNL 241 RQEDAFTPVR KIWEIFINQC RLNYVPGTNL TVDEQLLGFR GRCPFRMYIP NKPDKYGIKF 301 PMVCDAATKY MVDAIPYLGK STKTQGLPLG EFYVKELTQT VHGTNRNVTC DNWFTSVPLA 361 KSLLNSPYNL TLVGTIRSNK REIPEEVKNS RSRQVGSSMF CFDGPLTLVS YKPKPSKMVF 421 LLSSCNEDAV VNQSNGKPDM ILFYNQTKGG VDSFDQMCSS MSTNRKTNRW PMAVFYGMLN 481 MAFVNSYIIY CHNMLAKKEK PLSRKDFMKK LSTDLTTPSM QKRLEAPTLK RSLRDNITNV 541 LKIVPQAAID TSFDEPEPKK RRYCGFCSYK KKRMTKTQCF KCKKPVCGEH NIDVCQDCI (SEQ ID NO: 14534) . [0360] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Ctenoplusia agnata. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaaccctag aagcccaatc tacgtcattc tgacgtgtat gtcgccgaaa atactctgtc 61 tctttctcct gcacgatcgg attgccgcga acgctcgatt caacccagtt ggcgccgaga 121 tctattggag gactgcggcg ttgattcggt aagtcccgcc attttgtcat agtaacagta 181 ttgcacgtca gcttgacgta tatttgggct ttgtgttatt tttgtaaatt ttcaacgtta 241 gtttattatt gcatcttttt gttacattac tggtttattt gcatgtatta ctcaaatatt 301 atttttattt tagcgtagaa aataca (SEQ ID NO: 14535) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 agactgtttt ttttgtattt gcattatata ttatattcta aagttgattt aattctaaga 61 aaaacattaa aataagtttc tttttgtaaa atttaattaa ttataagaaa aagtttaagt 121 tgatctcatt ttttataaaa atttgcaatg tttccaaagt tattattgta aaagaataaa 181 taaaagtaaa ctgagtttta attgatgttt tattatatca ttatactata tattacttaa 241 ataaaacaat aactgaatgt atttctaaaa ggaatcacta gaaaatatag tgatcaaaaa 301 tttacacgtc atttttgcgt atgattgggc tttataggtt ctaaaaatat gattgggcct 361 ctagggttaa (SEQ ID NO: 14536) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCTAGAAGCCCAATC (SEQ ID NO: 14564).

[0361] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Agrotis ipsilon. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MESRQRLNQD EIATILENDD DYSPLDSDSE AEDRWEDDV WSDNEDAMID YVEDTSRQED

61 PDNNIASQES ANLEVTSLTS HRIISLPQRS ICGKNNHVWS TTKGRTTGRT SAINIIRTNR

121 GPTRMCRNIV DPLLCFQLFI TDEI IHEIVK WTNVEMIVKR QNLIDISASY RDTNTMEMWA

181 LVGILTLTAV MKDNHLSTDE LFDATFSGTR YVSVMSRERF EFLIRCMRMD DKTLRPTLRS

241 DDAFIPVRKL WEIFINQCRL NYVPGGNLTV DEQLLGFRGR CPFRMYIPNK PDKYGIRFPM

301 MCDAATKYMI DAIPYLGKST KTNGLPLGEF YVKELTKTVH GTNRNVTCDN WFTSIPLAKN

361 MLQAPYNLTI VGTIRSNKRE IPEEIKNSRS RPVGSSMFCF DGPLTLVSYK PKPSRMVFLL

421 SSCDENAVIN ESNGKPDMIL FYNQTKGGVD SFDQMCKSMS ANRKTNRWPM AVFYGMLNMA

481 FVNSYIIYCH NKINKQKKPI NRKEFMKNLS TDLTTPWMQE RLKAPTLKRT LRDNITNVLK

541 NWPPSPANN SEEPGPKKRS YCGFCSYKKR RMTKTQFYKC KKAICGEHNI DVCQDCV (SEQ ID NO: 14537) .

[0362] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Agrotis ipsilon. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaaccctag aagcccaatc tacgtaaatt tgacgtatac cgcggcgaaa tatatctgtc

61 tctttcacgt ttaccgtcgg attcccgcta acttcggaac caactcagta gccattgaga

121 actcccagga cacagttgcg tcatctcggt aagtgccgcc attttgttgt aatagacagg

181 ttgcacgtca ttttgacgta taattgggct ttgtgtaact tttgaaatta tttataattt

241 ttattgatgt gatttatttg agttaatcgt attgtttcgt tacatttttc atatgatatt

301 aatattttca gattgaatat aaa (SEQ ID NO: 14538) . In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of: 1 agactgtttt ttttaaaagg cttataaagt attactattg cgtgatttaa ttttataaaa 61 atatttaaaa ccagttgatt tttttaataa ttacctaatt ttaagaaaaa atgttagaag 121 cttgatattt ttgttgattt ttttctaaga tttgattaaa aggccataat tgtattaata 181 aagagtattt ttaacttcaa atttatttta tttattaatt aaaacttcaa ttatgataat 241 acatgcaaaa atatagttca tcaacagaaa aatataggaa aactctaata gttttatttt 301 tacacgtcat ttttacgtat gattgggctt tatagctagt caaatatgat tgggcttcta 361 gggttaa (SEQ ID NO: 14539) .

[0363] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Megachile rotundata. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MNGKDSLGEF YLDDLSDCLD CRSASSTDDE SDSSNIAIRK RCPIPLIYSD SEDEDMNNNV

61 EDNNHFVKES NRYHYQIVEK YKITSKTKKW KDVTVTEMKK FLGLI ILMGQ VKKDVLYDYW

121 STDPSIETPF FSKVMSRNRF LQIMQSWHFY NNNDISPNSH RLVKIQPVID YFKEKFNNVY

181 KSDQQLSLDE CLIPWRGRLS IKTYNPAKIT KYGILVRVLS EARTGYVSNF CVYAADGKKI

241 EETVLSVIGP YKNMWHHVYQ DNYYNSVNIA KIFLKNKLRV CGTIRKNRSL PQILQTVKLS

301 RGQHQFLRNG HTLLEVWNNG KRNVNMISTI HSAQMAESRN RSRTSDCPIQ KPISIIDYNK

361 YMKGVDRADQ YLSYYSIFRK TKKWTKRWM FFINCALFNS FKVYTTLNGQ KITYKNFLHK

421 AALSLIEDCG TEEQGTDLPN SEPTTTRTTS RVDHPGRLEN FGKHKLVNIV TSGQCKKPLR

481 QCRVCASKKK LSRTGFACKY CNVPLHKGDC FERYHSLKKY (SEQ ID NO: 14540) .

[0364] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Megachile rotundata. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaaataatg cccactctag atgaacttaa cactttaccg accggccgtc gattattcga 61 cgtttgctcc ccagcgctta ccgaccggcc atcgattatt cgacgtttgc ttcccagcgc 121 ttaccgaccg gtcatcgact tttgatcttt ccgttagatt tggttaggtc agattgacaa 181 gtagcaagca tttcgcattc tttattcaaa taatcggtgc ttttttctaa gctttagccc 241 ttagaa (SEQ ID NO: 14541) .

In certain embodiments, the the piggyBac or piggyBac-like transposon comprises a sequence of:

1 acaacttctt ttttcaacaa atattgttat atggattatt tatttattta tttatttatg

61 gtatatttta tgtttattta tttatggtta ttatggtata ttttatgtaa ataataaact

121 gaaaacgatt gtaatagatg aaataaatat tgttttaaca ctaatataat taaagtaaaa

181 gattttaata aatttcgtta ccctacaata acacgaagcg tacaatttta ccagagttta 241 ttaa (SEQ ID NO: 14542) .

[0365] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Bombus impatiens. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to: 1 MNEKNGIGEF YLDDLSDCPD SYSRSNSGDE SDGSDTI IRK RGSVLPPRYS DSEDDEINNV

61 EDNANNVENN DDIWSTNDEA IILEPFEGSP GLKIMPSSAE SVTDNV LFF GDDFFEHLVR

121 ESNRYHYQVM EKYKIPSKAK KWTDITVPEM KKFLGLIVLM GQIKKDVLYD YWSTDPSIET

181 PFFSQVMSRN RFVQIMQSWH FCNNDNIPHD SHRLAKIQPV IDYFRRKFND VYKPCQQLSL

241 DESIIPWRGR LSIKTYNPAK ITKYGILVRV LSEAVTGYVC NFDVYAADGK KLEDTAVIEP

301 YKNIWHQIYQ DNYYNSVKMA RILLKNKVRV CGTIRKNRGL PRSLKTIQLS RGQYEFRRNH

361 QILLEVWNNG RRNVNMISTI HSAQLMESRS KSKRSDVPIQ KPNSI IDYNK YMKGVDRADQ 421 YLAYYSIFRK TKKWTKRWM FFINCALFNS FRVYTILNGK NITYKNFLHK VAVSWIEDGE

481 TNCTEQDDNL PNSEPTRRAP RLDHPGRLSN YGKHKLINIV TSGRSLKPQR QCRVCAVQKK 541 RSRTCFVCKF CNVPLHKGDC FERYHTLKKY (SEQ ID NO: 14543) .

[0366] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Bombus impatiens. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaatttttt aacattttac cgaccgatag ccgattaatc gggtttttgc cgctgacgct 61 taccgaccga taacctatta atcggctttt tgtcgtcgaa gcttaccaac ctatagccta 121 cctatagtta atcggttgcc atggcgataa acaatctttc tcattatatg agcagtaatt 181 tgttatttag tactaaggta ccttgctcag ttgcgtcagt tgcgttgctt tgtaagctcc 241 cacagtttta taccaattcg aaaaacttac cgttcgcg (SEQ ID NO: 14544).

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 actatttcac atttgaacta aaaaccgttg taatagataa aataaatata atttagtatt

61 aatattatgg aaacaaaaga ttttattcaa tttaattatc ctatagtaac aaaaagcggc

121 caattttatc tgagcatacg aaaagcacag atactcccgc ccgacagtct aaaccgaaac

181 agagccggcg ccagggagaa tctgcgcctg agcagccggt cggacgtgcg tttgctgttg

241 aaccgctagt ggtcagtaaa ccagaaccag tcagtaagcc agtaactgat cagttaacta

301 gattgtatag ttcaaattga acttaatcta gtttttaagc gtttgaatgt tgtctaactt

361 cgttatatat tatattcttt ttaa (SEQ ID NO: 14545) .

[0367] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Mamestra brassicae. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MFSFVPNKEQ TRTVLIFCFH LKTTAAESHR PLVEAFGEQV PTVKTCERWF QRFKSGDFDV 61 DDKEHGKPPK RYEDAELQAL LDEDDAQTQK QLAEQLEVSQ QAVSNRLREG GKIQKVGR V 121 PHELNERQRE RRKNTCEILL SRYKRKSFLH RIVTGEEKWI FFVNPKRKKS YVDPGQPATS 181 TARPNRFGKK TRLCVWWDQS GVIYYELLKP GETVNTARYQ QQLINLNRAL QRKRPEYQKR 241 QHRVIFLHDN APSHTARAVR DTLETLNWEV LPHAAYSPDL APSDYHLFAS MGHALAEQRF 301 DSYESVEEWL DEWFAAKDDE FYWRGIHKLP ERWDNCVASD GKYFE (SEQ ID NO: 14546) .

[0368] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Mamestra brassicae. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttattgggtt gcccaaaaag taattgcgga tttttcatat acctgtcttt taaacgtaca 61 tagggatcga actcagtaaa actttgacct tgtgaaataa caaacttgac tgtccaacca 121 ccatagtttg gcgcgaattg agcgtcataa ttgttttgac tttttgcagt caac (SEQ

ID NO: 14547) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of: 1 atgatttttt ctttttaaac caattttaat tagttaattg atataaaaat ccgcaattac 61 tttttgggca acccaataa (SEQ ID NO: 14548) .

[0369] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Mayetiola destructor. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MENFENWRKR RHLREVLLGH FFAKKTAAES HRLLVEVYGE HALAKTQCFE WFQRFKSGDF 61 DTEDKERPGQ PKKFEDEELE ALLDEDCCQT QEELAKSLGV TQQAISKRLK AAGYIQKQGN 121 WVPHELKPRD VERRFCMSEM LLQRHKKKSF LSRIITGDEK WIHYDNSKRK KSYVKRGGRA 181 KSTPKSNLHG AKVMLCIWWD QRGVLYYELL EPGQTITGDL YRTQLIRLKQ ALAEKRPEYA 241 KRHGAVIFHH DNARPHVALP VKNYLENSGW EVLPHPPYSP DLAPSDYHLF RSMQNDLAGK 301 RFTSEQGIRK WLDSFLAAKP AKFFEKGIHE LSERWEKVIA SDGQYFE (SEQ ID NO: 14549) .

[0370] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Mayetiola destructor. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 taagacttcc aaaatttcca cccgaacttt accttccccg cgcattatgt ctctcttttc 61 accctctgat ccctggtatt gttgtcgagc acgatttata ttgggtgtac aacttaaaaa 121 ccggaattgg acgctagatg tccacactaa cgaatagtgt aaaagcacaa atttcatata 181 tacgtcattt tgaaggtaca tttgacagct atcaaaatca gtcaataaaa ctattctatc 241 tgtgtgcatc atattttttt attaact (SEQ ID NO: 14550) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tgcattcatt cattttgtta tcgaaataaa gcattaattt tcactaaaaa attccggttt 61 ttaagttgta cacccaatat catccttagt gacaattttc aaatggcttt cccattgagc 121 tgaaaccgtg gctctagtaa gaaaaacgcc caacccgtca tcatatgcct tttttttctc 181 aacatccg (SEQ ID NO: 14551) .

[0371] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Apis mellifera. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MENQKEHYRH ILLFYFRKGK NASQAHKKLC AVYGDEALKE RQCQNWFDKF RSGDFSLKDE

61 KRSGRPVEVD DDLIKAIIDS DRHSTTREIA EKLHVSHTCI ENHLKQLGYV QKLDTWVPHE

121 LKEKHLTQRI NSCDLLKKRN ENDPFLKRLI TGDEK WYN NIKRKRSWSR PREPAQTTSK

181 AGIHRKKVLL SVWWDYKGIV YFELLPPNRT INSWYIEQL TKLNNAVEEK RPELTNRKGV

241 VFHHDNARPH TSLVTRQKLL ELGWDVLPHP PYSPDLAPSD YFLFRSLQNS LNGKNFNNDD

301 DIKSYLIQFF ANKNQKFYER GIMMLPERWQ KVIDQNGQHI TE (SEQ ID NO: 14552) .

[0372] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Apis mellifera. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of: 1 ttgggttggc aactaagtaa ttgcggattt cactcataga tggcttcagt tgaattttta 61 ggtttgctgg cgtagtccaa atgtaaaaca cattttgtta tttgatagtt ggcaattcag 121 ctgtcaatca gtaaaaaaag ttttttgatc ggttgcgtag ttttcgtttg gcgttcgttg 181 aaaa (SEQ ID NO: 14553) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 agttatttag ttccatgaaa aaattgtctt tgattttcta aaaaaaatcc gcaattactt 61 agttgccaat ccaa (SEQ ID NO: 14554) .

[0373] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Messor bouvieri. The piggyBac

(PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,

70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MSSFVPENVH LRHALLFLFH QKKRAAESHR LLVETYGEHA PTIRTCETWF RQFKCGDFNV 61 QDKERPGRPK TFEDAELQEL LDEDSTQTQK QLAEKLNVSR VAICERLQAM GKIQKMGR V 121 PHELNDRQME NRKIVSEMLL QRYERKSFLH RIVTGDEKWI YFENPKRKKS WLSPGEAGPS 181 TARPNRFGRK TMLCVWWDQI GWYYELLKP GETVNTDRYR QQMINLNCAL IEKRPQYAQR 241 HDKVILQHDN APSHTAKPVK EMLKSLGWEV LSHPPYSPDL APSDYHLFAS MGHALAEQHF 301 ADFEEVKKWL DEWFSSKEKL FFWNGIHKLS ERWTKCIESN GQYFE (SEQ ID NO: 14555) .

[0374] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Messor bouvieri. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 agtcagaaat gacacctcga tcgacgacta atcgacgtct aatcgacgtc gattttatgt 61 caacatgtta ccaggtgtgt cggtaattcc tttccggttt ttccggcaga tgtcactagc 121 cataagtatg aaatgttatg atttgataca tatgtcattt tattctactg acattaacct 181 taaaactaca caagttacgt tccgccaaaa taacagcgtt atagatttat aattttttga 241 aa (SEQ ID NO: 14556) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ataaatttga actatccatt ctaagtaacg tgttttcttt aacgaaaaaa ccggaaaaga 61 attaccgaca ctcctggtat gtcaacatgt tattttcgac attgaatcgc gtcgattcga 121 agtcgatcga ggtgtcattt ctgact (SEQ ID NO: 14557) .

[0375] In certain embodiments of the methods of the disclosure, the transposase enzyme is a piggyBac or piggyBac-like transposase enzyme. In certain embodiments, the piggyBac or piggyBac-like transposase enzyme is isolated or derived from Trichoplusia ni. The piggyBac (PB) or piggyBac-like transposase enzyme may comprise or consist of an amino acid sequence at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or any percentage in between identical to:

1 MGSSLDDEHI LSALLQSDDE LVGEDSDSEV SDHVSEDDVQ SDTEEAFIDE VHEVQPTSSG

61 SEILDEQNVI EQPGSSLASN RILTLPQRTI RGKNKHCWST SKSTRRSRVS ALNIVRSQRG

121 PTRMCRNIYD PLLCFKLFFT DEIISEIVKW TNAEISLKRR ESMTSATFRD TNEDEIYAFF

181 GILVMTAVRK DNHMSTDDLF DRSLSMVYVS VMSRDRFDFL IRCLRMDDKS IRPTLRENDV

241 FTPVRKIWDL FIHQCIQNYT PGAHLTIDEQ LLGFRGRCPF RVYIPNKPSK YGIKILMMCD

301 SGTKYMINGM PYLGRGTQTN GVPLGEYYVK ELSKPVHGSC RNITCDNWFT SIPLAKNLLQ 361 EPYKLTIVGT VRSNKREIPE VLKNSRSRPV GTSMFCFDGP LTLVSYKPKP AKMVYLLSSC 421 DEDASINEST GKPQMVMYYN QTKGGVDTLD QMCSVMTCSR KTNRWPMALL YGMINIACIN

481 SFIIYSHNVS SKGEKVQSRK KFMRNLYMSL TSSFMRKRLE APTLKRYLRD NISNILPKEV 541 PGTSDDSTEE PVMKKRTYCT YCPSKIRRKA NASCKKCKKV ICREHNIDMC QSCF (SEQ

ID NO: 14558) .

[0376] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Trichoplusia ni. In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ttaaccctag aaagatagtc tgcgtaaaat tgacgcatgc attcttgaaa tattgctctc 61 tctttctaaa tagcgcgaat ccgtcgctgt gcatttagga catctcagtc gccgcttgga 121 gctcccgtga ggcgtgcttg tcaatgcggt aagtgtcact gattttgaac tataacgacc 181 gcgtgagtca aaatgacgca tgattatctt ttacgtgact tttaagattt aactcatacg 241 ataattatat tgttatttca tgttctactt acgtgataac ttattatata tatattttct 301 tgttatagat ate (SEQ ID NO: 14559) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat 61 aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat 121 atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgegtcaatt 181 ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg gttaa (SEQ ID NO: 14560) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 ccctagaaag atagtctgeg taaaattgac geatgeatte ttgaaatatt gctctctctt 61 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc 121 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt 181 gagtcaaaat gaegcatgat tatcttttac gtgactttta agatttaact catacgataa 241 ttatattgtt atttcatgtt etacttaegt gataacttat tatatatata ttttcttgtt 301 atagatatc (SEQ ID NO: 14561) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat 61 aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat 121 atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgegtcaatt 181 ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg g (SEQ ID NO: 14562) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc 61 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt 121 gagtcaaaat gaegcatgat tatcttttac gtgactttta agatttaact catacgataa 181 ttatattgtt atttcatgtt etacttaegt gataacttat tatatatata ttttcttgtt 241 atagatatc (SEQ ID NO: 14609) .

In certain embodiments, the piggyBac or piggyBac-like transposon comprises a sequence of:

1 tttgttactt tatagaagaa attttgagtt tttgtttttt tttaataaat aaataaacat 61 aaataaattg tttgttgaat ttattattag tatgtaagtg taaatataat aaaacttaat 121 atctattcaa attaataaat aaacctcgat atacagaccg ataaaacaca tgegtcaatt 181 ttacgcatga ttatctttaa cgtacgtcac aatatgatta tctttctagg g (SEQ ID NO: 14610) .

[0377] In certain embodiments,the piggyBac or piggyBac-like transposon comprises SEQ

ID NO: 14561 and SEQ ID NO: 14562, and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14558. In certain embodiments,the piggyBac or piggyBac-like transposon comprises SEQ ID NO: 14609 and SEQ ID NO: 14610, and the piggyBac or piggyBac-like transposase comprises SEQ ID NO: 14558.

[0378] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Aphis gossypii. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCTTCCAGCGGGCGCGC (SEQ ID NO:

14565).

[0379] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Chilo suppressalis. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCAGATTAGCCT (SEQ ID NO: 14566).

[0380] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Heliothis virescens. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCTTAATTACTCGCG (SEQ ID NO: 14567).

[0381] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Pectinophora gossypiella. In certain embodiments, the piggyBac or piggyBac- like transposon comprises an ITR sequence of CCCTAGATAACTAAAC (SEQ ID NO: 14568).

[0382] In certain embodiments, the piggyBac or piggyBac-like transposon is isolated or derived from Anopheles stephensi. In certain embodiments, the piggyBac or piggyBac-like transposon comprises an ITR sequence of CCCTAGAAAGATA (SEQ ID NO: 14569).

Gene Editing

[0383] In various embodiments, nucleases that may be used as cutting enzymes include, but are not limited to, Cas9, transcription activator-like effector nucleases (TALENs) and zinc finger nucleases. In certain embodiments, the Cas9 is a catalytically inactive or "inactivated" Cas9 (dCas9). In certain embodiments, the Cas9 is a catalytically inactive or "inactivated" nuclease domain of Cas9. In certain embodiments, the dCas9 is encoded by a shorter sequence that is derived from a full length, catalytically inactivated, Cas9, referred to herein as a "small" dCas9 or dSaCas9.

[0384] In certain embodiments, the inactivated, small, Cas9 (dSaCas9) operatively-linked to an active nuclease. In certain embodiments, the disclosure provides a fusion protein comprising, consisting essentially of or consisting of a DNA binding domain and molecule nuclease, wherein the nuclease comprises a small, inactivated Cas9 (dSaCas9). In certain embodiments, the dSaCas9 of the disclosure comprises the mutations D10A and N580A (underlined and bolded) which inactivate the catalytic site. In certain embodiments, the dSaCas9 (isolated or derived from Staphylococcus aureus) of the disclosure comprises the amino acid sequence of:

1 MKRNYILGLA IGITSVGYGI IDYETRDVID AGVRLFKEAN VENNEGRRSK RGARRLKRRR

61 RHRIQRVKKL LFDYNLLTDH SELSGINPYE ARVKGLSQKL SEEEFSAALL HLAKRRGVHN

121 VNEVEEDTGN ELSTKEQISR NSKALEEKYV AELQLERLKK DGEVRGSINR FKTSDYVKEA

181 KQLLKVQKAY HQLDQSFIDT YIDLLETRRT YYEGPGEGSP FGWKDIKEWY EMLMGHCTYF

241 PEELRSVKYA YNADLYNALN DLNNLVITRD ENEKLEYYEK FQIIENVFKQ KKKPTLKQIA

301 KEILVNEEDI KGYRVTSTGK PEFTNLKVYH DIKDITARKE IIENAELLDQ IAKILTIYQS

361 SEDIQEELTN LNSELTQEEI EQISNLKGYT GTHNLSLKAI NLILDELWHT NDNQIAIFNR

421 LKLVPKKVDL SQQKEIPTTL VDDFILSPW KRSFIQSIKV INAI IKKYGL PNDIIIELAR

481 EKNSKDAQKM INEMQKRNRQ TNERIEEIIR TTGKENAKYL IEKIKLHDMQ EGKCLYSLEA

541 IPLEDLLNNP FNYEVDHI IP RSVSFDNSFN NKVLVKQEEA SKKGNRTPFQ YLSSSDSKIS

601 YETFKKHILN LAKGKGRISK TKKEYLLEER DINRFSVQKD FINRNLVDTR YATRGLMNLL

661 RSYFRV NLD VKVKSINGGF TSFLRRKWKF KKERNKGYKH HAEDALI IAN ADFIFKEWKK

721 LDKAKKVMEN QMFEEKQAES MPEIETEQEY KEIFITPHQI KHIKDFKDYK YSHRVDKKPN

781 RELINDTLYS TRKDDKGNTL IV NLNGLYD KDNDKLKKLI NKSPEKLLMY HHDPQTYQKL

841 KLIMEQYGDE KNPLYKYYEE TGNYLTKYSK KDNGPVIKKI KYYGNKLNAH LDITDDYPNS

901 RNKWKLSLK PYRFDVYLDN GVYKFVTVKN LDVIKKENYY EVNSKCYEEA KKLKKISNQA

961 EFIASFYNND LIKINGELYR VIGVNNDLLN RIEV MIDIT YREYLENMND KRPPRIIKTI

1021 ASKTQSIKKY STDILGNLYE VKSKKHPQII KKG (SEQ ID NO: 14497)

[0385] In certain embodiments of the gene editing systems of the disclosure, the dCas9 of the disclosure comprises a dCas9 isolated or derived from Streptococcus pyogenes. In certain embodiments, the dCas9 comprises a dCas9 with substitutions at positions 10 and 840 of the amino acid sequence of the dCas9 which inactivate the catalytic site. In certain embodiments, these substitutions are D10A and H840A. In certain embodiments, the amino acid sequence of the dCas9 (isolated or derived from Streptococcus pyogenes) comprises the sequence of:

1 XDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61 ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121 NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181 VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241 LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301 LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361 GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421 AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481 WDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541 SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI

601 IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG

661 RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL 721 HEHIANLAGS PAIKKGILQT VKWDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER

781 MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA

841 IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEWKKMK NYWRQLLNAK LITQRKFDNL

901 TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS

961 KLVSDFRKDF QFYKVREINN YHHAHDAYLN AWGTALIKK YPKLESEFVY GDYKVYDVRK

1021 MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF

1081 ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA

1141 YSVLWAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK

1201 YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE

1261 QHKHYLDEI I EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA

1321 PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD (SEQ ID NO:

14498).

[0386] In certain embodiments of the gene editing systems of the disclosure, the nuclease domain may comprise, consist essentially of or consist of a dCas9 or a dSaCas9 and a type IIS endonuclease. In certain embodiments of the disclosure, the nuclease domain may comprise, consist essentially of or consist of a dSaCas9 and a type IIS endonuclease, including, but not limited to, Acil, Mnll, Alwl, Bbvl, Bed, BceAI, BsmAI, BsmFI, BspCNI, Bsrl, BtsCI, Hgal, Hphl, HpyAV, Mboll, Myll, Plel, SfaNI, Acul, BciVI, BfuAI, BmgBI, Bmrl, Bpml, BpuEI, Bsal, BseRI, Bsgl, Bsml, BspMI, BsrBI, BsrBI, BsrDI, BtgZI, Btsl, Earl, Ecil, Mmel, NmeAIII, BbvCI, BpulOI, BspQI, Sapl, Bael, BsaXI, CspCI, Bfil, Mboll, Acc36I, Fokl or Clo051. In certain embodiments of the disclosure, the nuclease domain may comprise, consist essentially of or consist of a dSaCas9 and Clo051. An exemplary Clo051 nuclease domain may comprise, consist essentially of or consist of, the amino acid sequence of:

EGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLFEMKVLELLVNEYGFKGRH LGGSRKPDGIVYSTTLEDNFGIIVDTKAYSEGYSLPISQADEMERYVRENSNRDEEVN PNKWWENFSEEVKKYYFVFISGSFKGKFEEQLRRLSMTTGVNGSAVNVVNLLLGAE KIRSGEMTIEELERAMFNNSEFILKY (SEQ ID NO: 14503).

[0387] An exemplary dCas9-Clo051 nuclease domain may comprise, consist essentially of or consist of, the amino acid sequence of (Clo051 sequence underlined, linker bold italics, dCas9 {Staphylococcus pyogenes) sequence in italics):

MAPKKKRKVEGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLFEMKVLELL VNEYGFKGRHLGGSRKPDGIVYSTTLEDNFGIIVDTKAYSEGYSLPISQADEMERYVR ENSNRDEEVNPNKWWENFSEEVKKYYFVFISGSFKGKFEEQLRRLSMTTGVNGSAV NVVNLLLGAEIQRSGEMTIEELERAMFNNSEFILKYG'G'G'G'.SPi^y^/GL^/GrN^ G^ VITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRI CY

LQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHL RKKL

VDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQL VQTYNQLFEENPINA

SGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAE DAKLQ

LSKDTYDDDLDNLLAQIGDQYADLFIAAKNLSDAILLSDILRVNTEITKAPLSASMI KRYDE

HHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMD GTE

ELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKIL TFRIP

YYVGPLARGNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIEPMTNFDKNLPNEKV LP

KHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKE DYFK

KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFE DREMIE

ERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFAN RNF

MQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKWDELVKVM GR

HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEK LYLY

YLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEE

VVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQI

LDSRMNTKYDENDKLIREVKVITLKSKL VSDFRKDFQFYKVREINNYHHAHDAYLNA WGT

ALIKKYPKLESEFVYGDYKVYD VRKMIAKSEQEIGKA TAKYFFYSNIMNFFKTEITLANGEI

RKRPLIETNGETGEIVWDKGRDFA TVRKVLSMPQ VNIVKKTEVQTGGFSKESILPKRNSDK

LIARKKD WDPKKYGGFDSPTVA YSVL WAKVEKGKSKKLKSVKELLGITIMERSSFEKNPI

DFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFL YLAS

HYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSA YNKHRDKP

IREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYET RIDLSQL

GG JGSPKKKRKVSS (SEQ ID NO: 14654).

[0388] Gene editing compositions of the disclosure may comprise a nuclease protein or a nuclease domain thereof. In certain embodiments, the gene editing composition comprises a sequence encoding a nuclease protein or a sequence encoding a nuclease domain thereof. In certain embodiments, the sequence encoding a nuclease protein or the sequence encoding a nuclease domain thereof comprises a DNA sequence, an RNA sequence, or a combination thereof. In certain embodiments, the nuclease or the nuclease domain thereof comprises one or more of a CRISPR/Cas protein, a Transcription Activator-Like Effector Nuclease (TALEN), a Zinc Finger Nuclease (ZFN), and an endonuclease. In certain embodiments, the nuclease or the nuclease domain thereof comprises one or more of a nuclease-inactivated Cas (dCas) protein, a Transcription Activator-Like Effector Nuclease (TALEN), a Zinc Finger Nuclease (ZFN), and an endonuclease. In certain embodiments, the nuclease or the nuclease domain thereof comprises a nuclease-inactivated Cas (dCas) protein and an endonuclease. In certain embodiments, the nuclease or the nuclease domain thereof comprises a nuclease- inactivated Cas9 (dCas9) protein and an endonuclease, wherein the endonuclease comprises a Clo051 nuclease or a nuclease domain thereof. In certain embodiments, the gene editing composition comprises a fusion protein. In certain embodiments, the fusion protein comprises a nuclease-inactivated Cas9 (dCas9) protein and a Clo051 nuclease or a Clo051 nuclease domain. In certain embodiments, the gene editing composition further comprises a guide sequence. In certain embodiments, the guide sequence comprises an RNA sequence.

[0389] In certain embodiments, the gene editing composition comprises a fusion protein. In certain embodiments, the fusion protein comprises a nuclease-inactivated Cas9 (dCas9) protein and a Clo051 nuclease or a Clo051 nuclease domain. In certain embodiments, the gene editing composition further comprises a guide sequence. In certain embodiments, the guide sequence comprises an RNA sequence. In certain embodiments, the fusion protein comprises or consists of the amino acid sequence:

MAPKKKRKVEGIKSNISLLKDELRGQISHISHEYLSLIDLAFDSKQNRLFEMKVLELLV EYGFKGRHLGGSRKP DGIVYSTTLEDNFGIIVDTKAYSEGYSLPISQADEMERYVRENSNRDEEV PNKWWENFSEEVKKYYFVFISGSF KGKFEEQLRRLSMTTGV GSAVNW LLLGAEKIRSGEMTIEELERAMFNNSEFILKYGGGGSDKKYSIGLAIGT NSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSN EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDK ADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSD AILLSDILRVNTEIT KAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFY KFIKPILEKMDGTEE LLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRI PYYVGPLARGNSRFA WMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNEL TKVKYVTEGMRKPA FLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLL KI IKDKDFLDNEENE DILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDK QSGKTILDFLKSDGF ANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKWDELVK VMGRHKPENIVIEM ARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMY VDQELDINRLSDYDV DAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEWKKMKNYWRQLLNAKLITQRKFDN LTKAERGGLSELDK AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFY KVREINNYHHAHDAY LNAWGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLI ETNGETGEIVWDKGRDFATVRKVLSMPQV IVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT VAYSVLWAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLP KYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEI IEQISEFSKRVILADANLD KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLI HQSITGLYETRIDLS

QLGGDGSPKKKRKVSS (SEQ ID NO: 1 65 ). In certain embodiments, the fusion protein is encoded by a nucleic acid comprising or consisting of the sequence: 1 atggcaccaa agaagaaaag aaaagtggag ggcatcaagt caaacatcag cctgctgaaa 61 gacgaactgc ggggacagat tagtcacatc agtcacgagt acctgtcact gattgatctg 121 gccttcgaca gcaagcagaa tagactgttt gagatgaaag tgctggaact gctggtcaac 181 gagtatggct tcaagggcag acatctgggc gggtctagga aacctgacgg catcgtgtac 241 agtaccacac tggaagacaa cttcggaatc attgtcgata ccaaggctta ttccgagggc 301 tactctctgc caattagtca ggcagatgag atggaaaggt acgtgcgcga aaactcaaat 361 agggacgagg aagtcaaccc caataagtgg tgggagaatt tcagcgagga agtgaagaaa 421 tactacttcg tctttatctc aggcagcttc aaagggaagt ttgaggaaca gctgcggaga 481 ctgtccatga ctaccggggt gaacggatct gctgtcaacg tggtcaatct gctgctgggc 541 gcagaaaaga tcaggtccgg ggagatgaca attgaggaac tggaacgcgc catgttcaac 601 aattctgagt ttatcctgaa gtatggaggc gggggaagcg ataagaaata ctccatcgga 661 ctggccattg gcaccaattc cgtgggctgg gctgtcatca cagacgagta caaggtgcca 721 agcaagaagt tcaaggtcct ggggaacacc gatcgccaca gtatcaagaa aaatctgatt 781 ggagccctgc tgttcgactc aggcgagact gctgaagcaa cccgactgaa gcggactgct 841 aggcgccgat atacccggag aaaaaatcgg atctgctacc tgcaggaaat tttcagcaac 901 gagatggcca aggtggacga tagtttcttt caccgcctgg aggaatcatt cctggtggag 961 gaagataaga aacacgagcg gcatcccatc tttggcaaca ttgtggacga agtcgcttat 1021 cacgagaagt accctactat ctatcatctg aggaagaaac tggtggactc caccgataag 1081 gcagacctgc gcctgatcta tctggccctg gctcacatga tcaagttccg ggggcatttt 1141 ctgatcgagg gagatctgaa ccctgacaat tctgatgtgg acaagctgtt catccagctg 1201 gtccagacat acaatcagct gtttgaggaa aacccaatta atgcctcagg cgtggacgca 1261 aaggccatcc tgagcgccag actgtccaaa tctaggcgcc tggaaaacct gatcgctcag 1321 ctgccaggag agaagaaaaa cggcctgttt gggaatctga ttgcactgtc cctgggcctg 1381 acacccaact tcaagtctaa ttttgatctg gccgaggacg ctaagctgca gctgtccaaa 1441 gacacttatg acgatgacct ggataacctg ctggctcaga tcggcgatca gtacgcagac 1501 ctgttcctgg ccgctaagaa tctgagtgac gccatcctgc tgtcagatat tctgcgcgtg 1561 aacacagaga ttactaaggc cccactgagt gcttcaatga tcaaaagata tgacgagcac 1621 catcaggatc tgaccctgct gaaggctctg gtgaggcagc agctgcccga gaaatacaag 1681 gaaatcttct ttgatcagag caagaatgga tacgccggct atattgacgg cggggcttcc 1741 caggaggagt tctacaagtt catcaagccc attctggaaa agatggacgg caccgaggaa 1801 ctgctggtga agctgaatcg ggaggacctg ctgagaaaac agaggacatt tgataacgga 1861 agcatccctc accagattca tctgggcgaa ctgcacgcca tcctgcgacg gcaggaggac 1921 ttctacccat ttctgaagga taaccgcgag aaaatcgaaa agatcctgac cttcagaatc 1981 ccctactatg tggggcctct ggcacgggga aatagtagat ttgcctggat gacaagaaag 2041 tcagaggaaa ctatcacccc ctggaacttc gaggaagtgg tcgataaagg cgctagcgca 2101 cagtccttca ttgaaaggat gacaaatttt gacaagaacc tgccaaatga gaaggtgctg 2161 cccaaacaca gcctgctgta cgaatatttc acagtgtata acgagctgac taaagtgaag 2221 tacgtcaccg aagggatgcg caagcccgca ttcctgtccg gagagcagaa gaaagccatc 2281 gtggacctgc tgtttaagac aaatcggaaa gtgactgtca aacagctgaa ggaagactat 2341 ttcaagaaaa ttgagtgttt cgattcagtg gaaatcagcg gcgtcgagga caggtttaac 2401 gcctccctgg ggacctacca cgatctgctg aagatcatca aggataagga cttcctggac 2461 aacgaggaaa atgaggacat cctggaggac attgtgctga cactgactct gtttgaggat 2521 cgcgaaatga tcgaggaacg actgaagact tatgcccatc tgttcgatga caaagtgatg 2581 aagcagctga aaagaaggcg ctacaccgga tggggacgcc tgagccgaaa actgatcaat 2641 gggattagag acaagcagag cggaaaaact atcctggact ttctgaagtc cgatggcttc 2701 gccaacagga acttcatgca gctgattcac gatgactctc tgaccttcaa ggaggacatc 2761 cagaaagcac aggtgtctgg ccagggggac agtctgcacg agcatatcgc aaacctggcc 2821 ggcagccccg ccatcaagaa agggattctg cagaccgtga aggtggtgga cgaactggtc 2881 aaggtcatgg gacgacacaa acctgagaac atcgtgattg agatggcccg cgaaaatcag 2941 acaactcaga agggccagaa aaacagtcga gaacggatga agagaatcga ggaaggcatc 3001 aaggagctgg ggtcacagat cctgaaggag catcctgtgg aaaacactca gctgcagaat 3061 gagaaactgt atctgtacta tctgcagaat ggacgggata tgtacgtgga ccaggagctg 3121 gatattaaca gactgagtga ttatgacgtg gatgccatcg tccctcagag cttcctgaag 3181 gatgactcca ttgacaacaa ggtgctgacc aggtccgaca agaaccgcgg caaatcagat 3241 aatgtgccaa gcgaggaagt ggtcaagaaa atgaagaact actggaggca gctgctgaat 3301 gccaagctga tcacacagcg gaaatttgat aacctgacta aggcagaaag aggaggcctg 3361 tctgagctgg acaaggccgg cttcatcaag cggcagctgg tggagacaag acagatcact 3421 aagcacgtcg ctcagattct ggatagcaga atgaacacaa agtacgatga aaacgacaag 3481 ctgatcaggg aggtgaaagt cattactctg aaatccaagc tggtgtctga ctttagaaag 3541 gatttccagt tttataaagt cagggagatc aacaactacc accatgctca tgacgcatac 3601 ctgaacgcag tggtcgggac cgccctgatt aagaaatacc ccaagctgga gtccgagttc 3661 gtgtacggag actataaagt gtacgatgtc cggaagatga tcgccaaatc tgagcaggaa 3721 attggcaagg ccaccgctaa gtatttcttt tacagtaaca tcatgaattt ctttaagacc 3781 gaaatcacac tggcaaatgg ggagatcaga aaaaggcctc tgattgagac caacggggag 3841 acaggagaaa tcgtgtggga caagggaagg gattttgcta ccgtgcgcaa agtcctgtcc 3901 atgccccaag tgaatattgt caagaaaact gaagtgcaga ccgggggatt ctctaaggag 3961 agtattctgc ctaagcgaaa ctctgataaa ctgatcgccc ggaagaaaga ctgggacccc 4021 aagaagtatg gcgggttcga ctctccaaca gtggcttaca gtgtcctggt ggtcgcaaag 4081 gtggaaaagg ggaagtccaa gaaactgaag tctgtcaaag agctgctggg aatcactatt 4141 atggaacgca gctccttcga gaagaatcct atcgattttc tggaagccaa gggctataaa 4201 gaggtgaaga aagacctgat cattaagctg ccaaaatact cactgtttga gctggaaaac 4261 ggacgaaagc gaatgctggc aagcgccgga gaactgcaga agggcaatga gctggccctg 4321 ccctccaaat acgtgaactt cctgtatctg gctagccact acgagaaact gaaggggtcc 4381 cctgaggata acgaacagaa gcagctgttt gtggagcagc acaaacatta tctggacgag 4441 atcattgaac agatttcaga gttcagcaag agagtgatcc tggctgacgc aaatctggat 4501 aaagtcctga gcgcatacaa caagcaccga gacaaaccaa tccgggagca ggccgaaaat 4561 atcattcatc tgttcaccct gacaaacctg ggcgcccctg cagccttcaa gtattttgac 4621 accacaatcg atcggaagag atacacttct accaaagagg tgctggatgc taccctgatc 4681 caccagagta ttaccggcct gtatgagaca cgcatcgacc tgtcacagct gggaggcgat 4741 gggagcccca agaaaaagcg gaaggtgtct agttaa (SEQ ID NO: 14655) .

[0390] In certain embodiments, the gene editing composition comprises a fusion protein. In certain embodiments, the fusion protein comprises a nuclease-inactivated Cas9 (dCas9) protein and a Clo051 nuclease or a Clo051 nuclease domain. In certain embodiments, the gene editing composition further comprises a guide sequence. In certain embodiments, the guide sequence comprises an RNA sequence. In certain embodiments, the fusion protein comprises or consists of the amino acid sequence:

1 MPKKKRKVEG IKSNISLLKD ELRGQISHIS HEYLSLIDLA FDSKQNRLFE MKVLELLVNE

61 YGFKGRHLGG SRKPDGIVYS TTLEDNFGII VDTKAYSEGY SLPISQADEM ERYVRENSNR

121 DEEVNPNKWW ENFSEEVKKY YFVFISGSFK GKFEEQLRRL SMTTGVNGSA VNWNLLLGA

181 EKIRSGEMTI EELERAMFNN SEFILKYGGG GSDKKYSIGL AIGTNSVGWA VITDEYKVPS

241 KKFKVLGNTD RHSIKKNLIG ALLFDSGETA EATRLKRTAR RRYTRRKNRI CYLQEIFSNE 301 MAKVDDSFFH RLEESFLVEE DKKHERHPIF GNIVDEVAYH EKYPTIYHLR KKLVDSTDKA 361 DLRLIYLALA HMIKFRGHFL IEGDLNPDNS DVDKLFIQLV QTYNQLFEEN PINASGVDAK 421 AILSARLSKS RRLENLIAQL PGEKKNGLFG NLIALSLGLT PNFKSNFDLA EDAKLQLSKD 481 TYDDDLDNLL AQIGDQYADL FLAAKNLSDA ILLSDILRVN TEITKAPLSA SMIKRYDEHH 541 QDLTLLKALV RQQLPEKYKE IFFDQSKNGY AGYIDGGASQ EEFYKFIKPI LEKMDGTEEL 601 LVKLNREDLL RKQRTFDNGS IPHQIHLGEL HAILRRQEDF YPFLKDNREK IEKILTFRIP 661 YYVGPLARGN SRFAWMTRKS EETITPWNFE EWDKGASAQ SFIERMTNFD KNLPNEKVLP 721 KHSLLYEYFT VYNELTKVKY VTEGMRKPAF LSGEQKKAIV DLLFKTNRKV TVKQLKEDYF 781 KKIECFDSVE ISGVEDRFNA SLGTYHDLLK I IKDKDFLDN EENEDILEDI VLTLTLFEDR 841 EMIEERLKTY AHLFDDKVMK QLKRRRYTGW GRLSRKLING IRDKQSGKTI LDFLKSDGFA 901 NRNFMQLIHD DSLTFKEDIQ KAQVSGQGDS LHEHIANLAG SPAIKKGILQ TVKWDELVK 961 VMGRHKPENI VIEMARENQT TQKGQKNSRE RMKRIEEGIK ELGSQILKEH PVENTQLQNE 1021 KLYLYYLQNG RDMYVDQELD INRLSDYDVD AIVPQSFLKD DSIDNKVLTR SDKNRGKSDN 1081 VPSEEWKKM KNYWRQLLNA KLITQRKFDN LTKAERGGLS ELDKAGFIKR QLVETRQITK 1141 HVAQILDSRM NTKYDENDKL IREVKVITLK SKLVSDFRKD FQFYKVREIN NYHHAHDAYL 1201 NAWGTALIK KYPKLESEFV YGDYKVYDVR KMIAKSEQEI GKATAKYFFY SNIMNFFKTE 1261 ITLANGEIRK RPLIETNGET GEIVWDKGRD FATVRKVLSM PQVNIVKKTE VQTGGFSKES

1321 ILPKRNSDKL IARKKDWDPK KYGGFDSPTV AYSVLWAKV EKGKSKKLKS VKELLGITIM

1381 ERSSFEKNPI DFLEAKGYKE VKKDLI IKLP KYSLFELENG RKRMLASAGE LQKGNELALP

1441 SKYV FLYLA SHYEKLKGSP EDNEQKQLFV EQHKHYLDEI IEQISEFSKR VILADANLDK

1501 VLSAYNKHRD KPIREQAENI IHLFTLTNLG APAAFKYFDT TIDRKRYTST KEVLDATLIH

1561 QSITGLYETR IDLSQLGGDG SPKKKRKV (SEQ ID NO: 14656). In Certain embodiments, the fusion protein is encoded by a nucleic acid comprising or consisting of the sequence:

1 atgcctaaga agaagcggaa ggtggaaggc atcaaaagca acatctccct cctgaaagac 61 gaactccggg ggcagattag ccacattagt cacgaatacc tctccctcat cgacctggct 121 ttcgatagca agcagaacag gctctttgag atgaaagtgc tggaactgct cgtcaatgag 181 tacgggttca agggtcgaca cctcggcgga tctaggaaac cagacggcat cgtgtatagt 241 accacactgg aagacaactt tgggatcatt gtggatacca aggcatactc tgagggttat 301 agtctgccca tttcacaggc cgacgagatg gaacggtacg tgcgcgagaa ctcaaataga 361 gatgaggaag tcaaccctaa caagtggtgg gagaacttct ctgaggaagt gaagaaatac 421 tacttcgtct ttatcagcgg gtccttcaag ggtaaatttg aggaacagct caggagactg 481 agcatgacta ccggcgtgaa tggcagcgcc gtcaacgtgg tcaatctgct cctgggcgct 541 gaaaagattc ggagcggaga gatgaccatc gaagagctgg agagggcaat gtttaataat 601 agcgagttta tcctgaaata cggtggcggt ggatccgata aaaagtattc tattggttta 661 gccatcggca ctaattccgt tggatgggct gtcataaccg atgaatacaa agtaccttca 721 aagaaattta aggtgttggg gaacacagac cgtcattcga ttaaaaagaa tcttatcggt 781 gccctcctat tcgatagtgg cgaaacggca gaggcgactc gcctgaaacg aaccgctcgg 841 agaaggtata cacgtcgcaa gaaccgaata tgttacttac aagaaatttt tagcaatgag 901 atggccaaag ttgacgattc tttctttcac cgtttggaag agtccttcct tgtcgaagag 961 gacaagaaac atgaacggca ccccatcttt ggaaacatag tagatgaggt ggcatatcat 1021 gaaaagtacc caacgattta tcacctcaga aaaaagctag ttgactcaac tgataaagcg 1081 gacctgaggt taatctactt ggctcttgcc catatgataa agttccgtgg gcactttctc 1141 attgagggtg atctaaatcc ggacaactcg gatgtcgaca aactgttcat ccagttagta 1201 caaacctata atcagttgtt tgaagagaac cctataaatg caagtggcgt ggatgcgaag 1261 gctattctta gcgcccgcct ctctaaatcc cgacggctag aaaacctgat cgcacaatta 1321 cccggagaga agaaaaatgg gttgttcggt aaccttatag cgctctcact aggcctgaca 1381 ccaaatttta agtcgaactt cgacttagct gaagatgcca aattgcagct tagtaaggac 1441 acgtacgatg acgatctcga caatctactg gcacaaattg gagatcagta tgcggactta 1501 tttttggctg ccaaaaacct tagcgatgca atcctcctat ctgacatact gagagttaat 1561 actgagatta ccaaggcgcc gttatccgct tcaatgatca aaaggtacga tgaacatcac 1621 caagacttga cacttctcaa ggccctagtc cgtcagcaac tgcctgagaa atataaggaa 1681 atattctttg atcagtcgaa aaacgggtac gcaggttata ttgacggcgg agcgagtcaa 1741 gaggaattct acaagtttat caaacccata ttagagaaga tggatgggac ggaagagttg 1801 cttgtaaaac tcaatcgcga agatctactg cgaaagcagc ggactttcga caacggtagc 1861 attccacatc aaatccactt aggcgaattg catgctatac ttagaaggca ggaggatttt 1921 tatccgttcc tcaaagacaa tcgtgaaaag attgagaaaa tcctaacctt tcgcatacct 1981 tactatgtgg gacccctggc ccgagggaac tctcggttcg catggatgac aagaaagtcc 2041 gaagaaacga ttactccatg gaattttgag gaagttgtcg ataaaggtgc gtcagctcaa 2101 tcgttcatcg agaggatgac caactttgac aagaatttac cgaacgaaaa agtattgcct 2161 aagcacagtt tactttacga gtatttcaca gtgtacaatg aactcacgaa agttaagtat 2221 gtcactgagg gcatgcgtaa acccgccttt ctaagcggag aacagaagaa agcaatagta 2281 gatctgttat tcaagaccaa ccgcaaagtg acagttaagc aattgaaaga ggactacttt 2341 aagaaaattg aatgcttcga ttctgtcgag atctccgggg tagaagatcg atttaatgcg 2401 tcacttggta cgtatcatga cctcctaaag ataattaaag ataaggactt cctggataac 2461 gaagagaatg aagatatctt agaagatata gtgttgactc ttaccctctt tgaagatcgg 2521 gaaatgattg aggaaagact aaaaacatac gctcacctgt tcgacgataa ggttatgaaa 2581 cagttaaaga ggcgtcgcta tacgggctgg ggacgattgt cgcggaaact tatcaacggg 2641 ataagagaca agcaaagtgg taaaactatt ctcgattttc taaagagcga cggcttcgcc 2701 aataggaact ttatgcagct gatccatgat gactctttaa ccttcaaaga ggatatacaa 2761 aaggcacagg tttccggaca aggggactca ttgcacgaac atattgcgaa tcttgctggt 2821 tcgccagcca tcaaaaaggg catactccag acagtcaaag tagtggatga gctagttaag 2881 gtcatgggac gtcacaaacc ggaaaacatt gtaatcgaga tggcacgcga aaatcaaacg 2941 actcagaagg ggcaaaaaaa cagtcgagag cggatgaaga gaatagaaga gggtattaaa 3001 gaactgggca gccagatctt aaaggagcat cctgtggaaa atacccaatt gcagaacgag 3061 aaactttacc tctattacct acaaaatgga agggacatgt atgttgatca ggaactggac 3121 ataaaccgtt tatctgatta cgacgtcgat gccattgtac cccaatcctt tttgaaggac 3181 gattcaatcg acaataaagt gcttacacgc tcggataaga accgagggaa aagtgacaat 3241 gttccaagcg aggaagtcgt aaagaaaatg aagaactatt ggcggcagct cctaaatgcg 3301 aaactgataa cgcaaagaaa gttcgataac ttaactaaag ctgagagggg tggcttgtct 3361 gaacttgaca aggccggatt tattaaacgt cagctcgtgg aaacccgcca aatcacaaag 3421 catgttgcac agatactaga ttcccgaatg aatacgaaat acgacgagaa cgataagctg 3481 attcgggaag tcaaagtaat cactttaaag tcaaaattgg tgtcggactt cagaaaggat 3541 tttcaattct ataaagttag ggagataaat aactaccacc atgcgcacga cgcttatctt 3601 aatgccgtcg tagggaccgc actcattaag aaatacccga agctagaaag tgagtttgtg 3661 tatggtgatt acaaagttta tgacgtccgt aagatgatcg cgaaaagcga acaggagata 3721 ggcaaggcta cagccaaata cttcttttat tctaacatta tgaatttctt taagacggaa 3781 atcactctgg caaacggaga gatacgcaaa cgacctttaa ttgaaaccaa tggggagaca 3841 ggtgaaatcg tatgggataa gggccgggac ttcgcgacgg tgagaaaagt tttgtccatg 3901 ccccaagtca acatagtaaa gaaaactgag gtgcagaccg gagggttttc aaaggaatcg 3961 attcttccaa aaaggaatag tgataagctc atcgctcgta aaaaggactg ggacccgaaa 4021 aagtacggtg gcttcgatag ccctacagtt gcctattctg tcctagtagt ggcaaaagtt 4081 gagaagggaa aatccaagaa actgaagtca gtcaaagaat tattggggat aacgattatg 4141 gagcgctcgt cttttgaaaa gaaccccatc gacttccttg aggcgaaagg ttacaaggaa 4201 gtaaaaaagg atctcataat taaactacca aagtatagtc tgtttgagtt agaaaatggc 4261 cgaaaacgga tgttggctag cgccggagag cttcaaaagg ggaacgaact cgcactaccg 4321 tctaaatacg tgaatttcct gtatttagcg tcccattacg agaagttgaa aggttcacct 4381 gaagataacg aacagaagca actttttgtt gagcagcaca aacattatct cgacgaaatc 4441 atagagcaaa tttcggaatt cagtaagaga gtcatcctag ctgatgccaa tctggacaaa 4501 gtattaagcg catacaacaa gcacagggat aaacccatac gtgagcaggc ggaaaatatt 4561 atccatttgt ttactcttac caacctcggc gctccagccg cattcaagta ttttgacaca 4621 acgatagatc gcaaacgata cacttctacc aaggaggtgc tagacgcgac actgattcac 4681 caatccatca cgggattata tgaaactcgg atagatttgt cacagcttgg gggtgacgga 4741 tcccccaaga agaagaggaa agtctga (SEQ ID NO: 14657) .

[0391] In certain embodiments, the dCas9 of the disclosure comprises a dCas9 isolated or derived from Staphyloccocus pyogenes. In certain embodiments, the dCas9 comprises a dCas9 with substitutions at positions 10 and 840 of the amino acid sequence of the dCas9, which inactivate the catalytic site. In certain embodiments, these substitutions are DIOA and H840A. In certain embodiments, the "X" residue at position 1 of the dCas9 sequence is a methionine (M). In certain embodiments, the amino acid sequence of the dCas9 comprises the sequence of:

1 XDKKYSIGLA IGTNSVGWAV ITDEYKVPSK KFKVLGNTDR HSIKKNLIGA LLFDSGETAE

61 ATRLKRTARR RYTRRKNRIC YLQEIFSNEM AKVDDSFFHR LEESFLVEED KKHERHPIFG

121 NIVDEVAYHE KYPTIYHLRK KLVDSTDKAD LRLIYLALAH MIKFRGHFLI EGDLNPDNSD

181 VDKLFIQLVQ TYNQLFEENP INASGVDAKA ILSARLSKSR RLENLIAQLP GEKKNGLFGN

241 LIALSLGLTP NFKSNFDLAE DAKLQLSKDT YDDDLDNLLA QIGDQYADLF LAAKNLSDAI

301 LLSDILRVNT EITKAPLSAS MIKRYDEHHQ DLTLLKALVR QQLPEKYKEI FFDQSKNGYA

361 GYIDGGASQE EFYKFIKPIL EKMDGTEELL VKLNREDLLR KQRTFDNGSI PHQIHLGELH

421 AILRRQEDFY PFLKDNREKI EKILTFRIPY YVGPLARGNS RFAWMTRKSE ETITPWNFEE

481 WDKGASAQS FIERMTNFDK NLPNEKVLPK HSLLYEYFTV YNELTKVKYV TEGMRKPAFL

541 SGEQKKAIVD LLFKTNRKVT VKQLKEDYFK KIECFDSVEI SGVEDRFNAS LGTYHDLLKI 601 IKDKDFLDNE ENEDILEDIV LTLTLFEDRE MIEERLKTYA HLFDDKVMKQ LKRRRYTGWG 661 RLSRKLINGI RDKQSGKTIL DFLKSDGFAN RNFMQLIHDD SLTFKEDIQK AQVSGQGDSL 721 HEHIANLAGS PAIKKGILQT VKWDELVKV MGRHKPENIV IEMARENQTT QKGQKNSRER 781 MKRIEEGIKE LGSQILKEHP VENTQLQNEK LYLYYLQNGR DMYVDQELDI NRLSDYDVDA 841 IVPQSFLKDD SIDNKVLTRS DKNRGKSDNV PSEEWKKMK NYWRQLLNAK LITQRKFDNL 901 TKAERGGLSE LDKAGFIKRQ LVETRQITKH VAQILDSRMN TKYDENDKLI REVKVITLKS 961 KLVSDFRKDF QFYKVREINN YHHAHDAYLN AWGTALIKK YPKLESEFVY GDYKVYDVRK 1021 MIAKSEQEIG KATAKYFFYS NIMNFFKTEI TLANGEIRKR PLIETNGETG EIVWDKGRDF 1081 ATVRKVLSMP QVNIVKKTEV QTGGFSKESI LPKRNSDKLI ARKKDWDPKK YGGFDSPTVA 1141 YSVLWAKVE KGKSKKLKSV KELLGITIME RSSFEKNPID FLEAKGYKEV KKDLIIKLPK 1201 YSLFELENGR KRMLASAGEL QKGNELALPS KYVNFLYLAS HYEKLKGSPE DNEQKQLFVE 1261 QHKHYLDEI I EQISEFSKRV ILADANLDKV LSAYNKHRDK PIREQAENII HLFTLTNLGA 1321 PAAFKYFDTT IDRKRYTSTK EVLDATLIHQ SITGLYETRI DLSQLGGD (SEQ ID NO: 14498) .

[0392] In certain embodiments, the dCas9 of the disclosure comprises a dCas9 isolated or derived from Staphylococcus aureus. In certain embodiments, the dCas9 comprises a dCas9 with substitutions at positions 10 and 580 of the amino acid sequence of the dCas9 which inactivate the catalytic site. In certain embodiments, these substitutions are D10A and N580A. In certain embodiments, the dCas9 is a small and inactive Cas9 (dSaCas9). In certain embodiments, the amino acid sequence of the dSaCas9 comprises the sequence of:

1 mkrnyilglA igitsvgygi idyetrdvid agvrlfkean vennegrrsk rgarrlkrrr

61 rhriqrvkkl lfdynlltdh selsginpye arvkglsqkl seeefsaall hlakrrgvhn

121 vneveedtgn elstkeqisr nskaleekyv aelqlerlkk dgevrgsinr fktsdyvkea

181 kqllkvqkay hqldqsfidt yidlletrrt yyegpgegsp fgwkdikewy emlmghctyf

241 peelrsvkya ynadlynaln dlnnlvitrd enekleyyek fqiienvfkq kkkptlkqia

301 keilvneedi kgyrvtstgk peftnlkvyh dikditarke iienaelldq iakiltiyqs

361 sediqeeltn lnseltqeei eqisnlkgyt gthnlslkai nlildelwht ndnqiaifnr

421 lklvpkkvdl sqqkeipttl vddfilsp v krsfiqsikv inaiikkygl pndiiielar

481 eknskdaqkm inemqkrnrq tnerieeiir ttgkenakyl iekiklhdmq egkclyslea

541 ipledllnnp fnyevdhiip rsvsfdnsfn nkvlvkqeeA skkgnrtpfq ylsssdskis

601 yetfkkhiln lakgkgrisk tkkeylleer dinrfsvqkd finrnlvdtr yatrglmnll

661 rsyfrvnnld vkvksinggf tsflrrkwkf kkernkgykh haedaliian adfifkewkk

721 ldkakkvmen qmfeekqaes mpeieteqey keifitphqi khikdfkdyk yshrvdkkpn

781 relindtlys trkddkgntl ivnnlnglyd kdndklkkli nkspekllmy hhdpqtyqkl

841 klimeqygde knplykyyee tgnyltkysk kdngpvikki kyygnklnah lditddypns

901 rnk vklslk pyrfdvyldn gvykfvtvkn ldvikkenyy evnskcyeea kklkkisnqa

961 efiasfynnd likingelyr vigvnndlln rievnmidit yreylenmnd krppriikti

1021 asktqsikky stdilgnlye vkskkhpqii kkg (SEQ ID NO: 14658) . [0393] In certain embodiments of the gene editing systems described herein, the nuclease may comprise, consist essentially of or consist of, a homodimer or a heterodimer. Nuclease domains of the disclosure may comprise, consist essentially of or consist of a nuclease domain isolated, derived or recombined from a transcription-activator-like effector nuclease (TALEN). TALENs are transcription factors with programmable DNA binding domains that provide a means to create designer proteins that bind to pre-determined DNA sequences or individual nucleic acids. Modular DNA binding domains have been identified in

transcriptional activator-like (TAL) proteins, or, more specifically, transcriptional activatorlike effector nucleases (TALENs), thereby allowing for the de novo creation of synthetic transcription factors that bind to DNA sequences of interest and, if desirable, also allowing a second domain present on the protein or polypeptide to perform an activity related to DNA. TAL proteins have been derived from the organisms Xanthomonas and Ralstonia.

[0394] In certain embodiments of the gene editing systems described herein, the nuclease domain may comprise, consist essentially of or consist of a nuclease domain isolated, derived or recombined from a TALEN and a type IIS endonuclease. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of Acil, Mnll, Alwl, Bbvl, Bed, BceAI, BsmAI, BsmFI, BspCNI, BsrI, BtsCI, Hgal, HphI, HpyAV, Mboll, Myll, Plel, SfaNI, Acul, BciVI, BfuAI, BmgBI, Bmrl, Bpml, BpuEI, Bsal, BseRI, Bsgl, Bsml, BspMI, BsrBI, BsrBI, BsrDI, BtgZI, Btsl, Earl, Ecil, Mmel, NmeAIII, BbvCI, BpulOI, BspQI, Sapl, Bael, BsaXI, CspCI, Bfil, Mboll, Acc36I, Fokl or Clo051. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of Clo051 (SEQ ID NO: 14503).

[0395] In certain embodiments of the gene editing systems described herein, the nuclease domain of may comprise, consist essentially of or consist of a nuclease domain isolated, derived or recombined from a zinc finger nuclease (ZFN) and a type IIS endonuclease. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of Acil, Mnll, Alwl, Bbvl, Bed, BceAI, BsmAI, BsmFI, BspCNI, BsrI, BtsCI, Hgal, HphI, HpyAV, Mboll, Myll, Plel, SfaNI, Acul, BciVI, BfuAI, BmgBI, Bmrl, Bpml, BpuEI, Bsal, BseRI, Bsgl, Bsml, BspMI, BsrBI, BsrBI, BsrDI, BtgZI, Btsl, Earl, Ecil, Mmel, NmeAIII, BbvCI, BpulOI, BspQI, Sapl, Bael, BsaXI, CspCI, Bfil, Mboll, Acc36I, Fokl or Clo051. In certain embodiments of the disclosure, the type IIS endonuclease may comprise, consist essentially of or consist of Clo051 (SEQ ID NO: 14503).

[0396] In certain embodiments of the gene editing systems described herein, the DNA binding domain and the nuclease domain may be covalently linked. For example, a fusion protein may comprise the DNA binding domain and the nuclease domain. In certain embodiments of the genomic editing compositions or constructs of the disclosure, the DNA binding domain and the nuclease domain may be operably linked through a non-covalent linkage.

Therapeutic Proteins

[0397] In certain embodiments of the composition and methods of the disclosure, modified immune or immune precursor cells express therapeutic proteins. Therapeutic proteins of the disclosure include secreted proteins. Preferably, in a therapeutic context, the therapeutic protein is a human protein, including a secreted human protein. When expressed or secreted by immune or immune precursor cells of the disclosure, the combination comprising the immune or immune precursor cell and the therapeutic protein secreted therefrom may be considered a monotherapy. However, the immune or immune precursor cells of the disclosure may be administered as a combination therapy with a second agent. Human therapeutic proteins of the disclosure include, but are not limited to, those provided at Table 1.

[0398] Table 1. Exemplary Human Secreted Proteins

CTD- SEQID NOS: 81-84

2370N5.3

RP11- SEQID NOS: 85-87 196G11.1

AC136352.5 SEQID NO: 88

RP11- SEQID NO: 89 812E19.9

AC145212.4 MaFF-interacting protein SEQID NO: 90

AC233755.1 SEQID NO: 91

AC011513.3 SEQID NOS: 92-93

ACACB Acetyl-CoA carboxylase beta SEQID NOS: 94- 100

ACAN Aggrecan SEQID NOS: 101- 108

ACE Angiotensin 1 converting enzyme SEQID NOS: 109- 121

ACHE Acetylcholinesterase (Yt blood group) SEQID NOS: 122- 134

ACP2 Acid phosphatase 2, lysosomal SEQID NOS: 135- 142

ACP5 Acid phosphatase 5, tartrate resistant SEQID NOS: 143- 151

ACP6 Acid phosphatase 6, lysophosphatidic SEQID NOS: 152- 158

PAPL Iron/zinc purple acid phosphatase-like protein SEQID NOS: 159- 162

ACPP Acid phosphatase, prostate SEQID NOS: 163- 167

ACR Acrosin SEQID NOS: 168- 169

ACRBP Acrosin binding protein SEQID NOS: 170- 174

ACRV1 Acrosomal vesicle protein 1 SEQID NOS: 175- 178

ACSF2 Acyl-CoA synthetase family member 2 SEQID NOS: 179- 187

ACTL10 Actin-like 10 SEQID NO: 188

ACVR1 Activin A receptor, type 1 SEQID NOS: 189- 197

ACVR1C Activin A receptor, type IC SEQID NOS: 198-201

ACVRL1 Activin A receptor type ll-like 1 SEQID NOS: 202-207

ACYP1 Acylphosphatase 1, erythrocyte (common) type SEQID NOS: 208-213

ACYP2 Acylphosphatase 2, muscle type SEQID NOS: 214-221

CECR1 Cat eye syndrome chromosome region, candidate 1 SEQID NOS: 222-229

ADAM10 ADAM metallopeptidase domain 10 SEQID NOS: 230-237

ADAM12 ADAM metallopeptidase domain 12 SEQID NOS: 238-240

ADAM15 ADAM metallopeptidase domain 15 SEQID NOS: 241-252

ADAM17 ADAM metallopeptidase domain 17 SEQID NOS: 253-255

ADAM18 ADAM metallopeptidase domain 18 SEQID NOS: 256-260

ADAM22 ADAM metallopeptidase domain 22 SEQID NOS: 261-269

ADAM28 ADAM metallopeptidase domain 28 SEQID NOS: 270-275

ADAM29 ADAM metallopeptidase domain 29 SEQID NOS: 276-284

ADAM32 ADAM metallopeptidase domain 32 SEQID NOS: 285-291

ADAM33 ADAM metallopeptidase domain 33 SEQID NOS: 292-296

ADAM7 ADAM metallopeptidase domain 7 SEQID NOS: 297-300

ADAM8 ADAM metallopeptidase domain 8 SEQID NOS: 301-305

ADAM9 ADAM metallopeptidase domain 9 SEQID NOS: 306-311

ADAMDEC1 ADAM-like, decysin 1 SEQID NOS: 312-314

ADAMTS1 ADAM metallopeptidase with thrombospondin type SEQID NOS: 315-318

1 motif, 1 ADAMTS10 ADAM metallopeptidase with thrombospondin type SEQID NOS: 319-324 1 motif, 10

ADAMTS12 ADAM metallopeptidase with thrombospondin type SEQID NOS: 325-327

1 motif, 12

ADAMTS13 ADAM metallopeptidase with thrombospondin type SEQID NOS: 328-335

1 motif, 13

ADAMTS14 ADAM metallopeptidase with thrombospondin type SEQID NOS: 336-337

1 motif, 14

ADAMTS15 ADAM metallopeptidase with thrombospondin type SEQID NO: 338

1 motif, 15

ADAMTS16 ADAM metallopeptidase with thrombospondin type SEQID NOS: 339-340

1 motif, 16

ADAMTS17 ADAM metallopeptidase with thrombospondin type SEQID NOS: 341-344

1 motif, 17

ADAMTS18 ADAM metallopeptidase with thrombospondin type SEQID NOS: 345-348

1 motif, 18

ADAMTS19 ADAM metallopeptidase with thrombospondin type SEQID NOS: 349-352

1 motif, 19

ADAMTS2 ADAM metallopeptidase with thrombospondin type SEQID NOS: 353-355

1 motif, 2

ADAMTS20 ADAM metallopeptidase with thrombospondin type SEQID NOS: 356-359

1 motif, 20

ADAMTS3 ADAM metallopeptidase with thrombospondin type SEQID NOS: 360-361

1 motif, 3

ADAMTS5 ADAM metallopeptidase with thrombospondin type SEQID NO: 362

1 motif, 5

ADAMTS6 ADAM metallopeptidase with thrombospondin type SEQID NOS: 363-364

1 motif, 6

ADAMTS7 ADAM metallopeptidase with thrombospondin type SEQID NO: 365

1 motif, 7

ADAMTS8 ADAM metallopeptidase with thrombospondin type SEQID NO: 366

1 motif, 8

ADAMTS9 ADAM metallopeptidase with thrombospondin type SEQID NOS: 367-371

1 motif, 9

ADAMTSL1 ADAMTS-like 1 SEQID NOS: 372-382

ADAMTSL2 ADAMTS-like 2 SEQID NOS: 383-385

ADAMTSL3 ADAMTS-like 3 SEQID NOS: 386-387

ADAMTSL4 ADAMTS-like 4 SEQID NOS: 388-391

ADAMTSL5 ADAMTS-like 5 SEQID NOS: 392-397

ADCK1 AarF domain containing kinase 1 SEQID NOS: 398-402

ADCYAP1 Adenylate cyclase activating polypeptide 1 SEQID NOS: 403-404

(pituitary)

ADCYAP1R1 Adenylate cyclase activating polypeptide 1 SEQID NOS: 405-411

(pituitary) receptor type 1

ADGRA3 Adhesion G protein-coupled receptor A3 SEQID NOS: 412-416

ADGRB2 Adhesion G protein-coupled receptor B2 SEQID NOS: 417-425

ADGRD1 Adhesion G protein-coupled receptor Dl SEQID NOS: 426-431

ADGRE3 Adhesion G protein-coupled receptor E3 SEQID NOS: 432-436

ADGRE5 Adhesion G protein-coupled receptor E5 SEQID NOS: 437-442 ADGRF1 Adhesion G protein-coupled receptor Fl SEQID NOS: 443-447

ADGRG1 Adhesion G protein-coupled receptor Gl SEQID NOS: 448-512

ADGRG5 Adhesion G protein-coupled receptor G5 SEQID NOS: 513-515

ADGRG6 Adhesion G protein-coupled receptor G6 SEQID NOS: 516-523

ADGRV1 Adhesion G protein-coupled receptor VI SEQID NOS: 524-540

ADI1 Acireductone dioxygenase 1 SEQID NOS: 541-543

ADIG Adipogenin SEQID NOS: 544-547

ADIPOQ Adiponectin, ClQand collagen domain containing SEQID NOS: 548-549

ADM Adrenomedullin SEQID NOS: 550-557

ADM2 Adrenomedullin 2 SEQID NOS: 558-559

ADM5 Adrenomedullin 5 (putative) SEQID NO: 560

ADPGK ADP-dependent glucokinase SEQID NOS: 561-570

ADPRHL2 ADP-ribosylhydrolase like 2 SEQID NO: 571

AEBP1 AE binding protein 1 SEQID NOS: 572-579

LAC El Lactation elevated 1 SEQID NOS: 580-583

AFM Afamin SEQID NO: 584

AFP Alpha-fetoprotein SEQID NOS: 585-586

AGA Aspartylglucosaminidase SEQID NOS: 587-589

AGER Advanced glycosylation end product-specific SEQID NOS: 590-600 receptor

AGK Acylglycerol kinase SEQID NOS: 601-606

AG PS Alkylglycerone phosphate synthase SEQID NOS: 607-610

AGR2 Anterior gradient 2, protein disulphide isomerase SEQID NOS: 611-614 family member

AGR3 Anterior gradient 3, protein disulphide isomerase SEQID NOS: 615-617 family member

AGRN Agrin SEQID NOS: 618-621

AGRP Agouti related neuropeptide SEQID NO: 622

AGT Angiotensinogen (serpin peptidase inhibitor, clade A, SEQID NO: 623 member 8)

AGTPBP1 ATP/GTP binding protein 1 SEQID NOS: 624-627

AGTRAP Angiotensin II receptor-associated protein SEQID NOS: 628-635

AHCYL2 Adenosylhomocysteinase-like 2 SEQID NOS: 636-642

AHSG Alpha-2-HS-glycoprotein SEQID NOS: 643-644

AIG1 Androgen-induced 1 SEQID NOS: 645-653

AK4 Adenylate kinase 4 SEQID NOS: 654-657

AKAP10 A kinase (PRKA) anchor protein 10 SEQID NOS: 658-666

AKR1C1 Aldo-keto reductase family 1, member CI SEQID NOS: 667-669

RP4- SEQID NOS: 670-672

576H24.4

SERPINA3 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NO: 673 antiproteinase, antitrypsin), member 3

RP11-14J7.7 SEQID NOS: 674-675

RP11- SEQID NO: 676 903H12.5

AL356289.1 SEQID NO: 677

AL589743.1 SEQID NO: 678 XXbac- SEQID NOS: 679-680

BPG116M5.

17

XXbac- SEQID NO: 681

BPG181M17

.5

XXbac- SEQID NO: 682 BPG32J3.20

RP11- SEQID NO: 683 350014.18

A LAS 2 5'-aminolevulinate synthase 2 SEQID NOS: 684-691

ALB Albumin SEQID NOS: 692-701

ALDH9A1 Aldehyde dehydrogenase 9 family, member Al SEQID NO: 702

ALDOA Aldolase A, fructose-bisphosphate SEQID NOS: 703-717

ALG1 ALG1, chitobiosyldiphosphodolichol beta- SEQID NOS: 718-723 man nosyltransferase

ALG5 ALG5, dolichyl-phosphate beta-glucosyltransferase SEQID NOS: 724-725

ALG9 ALG9, a lpha-l,2-man nosyltransferase SEQID NOS: 726-736

FAM150A Family with sequence similarity 150, member A SEQID NOS: 737-738

FAM150B Family with sequence similarity 150, member B SEQID NOS: 739-745

ALKBH1 AlkB homolog 1, histone H2A dioxygenase SEQID NOS: 746-748

ALKBH5 AlkB homolog 5, RNA demethylase SEQID NOS: 749-750

ALPI Alkaline phosphatase, intestinal SEQID NOS: 751-752

ALPL Alkaline phosphatase, liver/bone/kidney SEQID NOS: 753-757

ALPP Alkaline phosphatase, placental SEQID NO: 758

ALPPL2 Alkaline phosphatase, placental-like 2 SEQID NO: 759

AMBN Ameloblastin (enamel matrix protein) SEQID NOS: 760-762

AMBP Alpha-l-microglobulin/bikunin precursor SEQID NOS: 763-765

AMELX Amelogenin, X-linked SEQID NOS: 766-768

AM ELY Amelogenin, Y-linked SEQID NOS: 769-770

AMH Anti-Mullerian hormone SEQID NO: 771

AMPD1 Adenosine monophosphate deaminase 1 SEQID NOS: 772-774

AMTN Amelotin SEQID NOS: 775-776

AMY1A Amylase, alpha 1A (salivary) SEQID NOS: 777-779

AMY1B Amylase, alpha IB (salivary) SEQID NOS: 780-783

AMY1C Amylase, alpha 1C (salivary) SEQID NO: 784

AMY2A Amylase, alpha 2A (pancreatic) SEQID NOS: 785-787

AMY2B Amylase, alpha 2B (pancreatic) SEQID NOS: 788-792

ANG Angiogenin, ribonuclease, RNase A family, 5 SEQID NOS: 793-794

ANGEL1 Angel homolog 1 (Drosophila) SEQID NOS: 795-798

ANGPT1 Angiopoietin 1 SEQID NOS: 799-803

ANGPT2 Angiopoietin 2 SEQID NOS: 804-807

ANGPT4 Angiopoietin 4 SEQID NO: 808

ANGPTL1 Angiopoietin-like 1 SEQID NOS: 809-811

ANGPTL2 Angiopoietin-like 2 SEQID NOS: 812-813

ANGPTL3 Angiopoietin-like 3 SEQID NO: 814

ANGPTL4 Angiopoietin-like 4 SEQID NOS: 815-822 ANGPTL5 Angiopoietin-like 5 SEQID NOS: 823-824

ANGPTL6 Angiopoietin-like 6 SEQID NOS: 825-827

ANGPTL7 Angiopoietin-like 7 SEQID NO: 828

C19orf80 Chromosome 19 open reading frame 80 SEQID NOS: 829-832

ANK1 Ankyrin 1, erythrocytic SEQID NOS: 833-843

ANKDD1A Ankyrin repeat and death domain containing 1A SEQID NOS: 844-850

ANKRD54 Ankyrin repeat domain 54 SEQID NOS: 851-859

ANKRD60 Ankyrin repeat domain 60 SEQID NO: 860

AN07 Anoctamin 7 SEQID NOS: 861-864

ANOS1 Anosmin 1 SEQID NO: 865

ANTXR1 Anthrax toxin receptor 1 SEQID NOS: 866-869

AOAH Acyloxyacyl hydrolase (neutrophil) SEQID NOS: 870-874

AOC1 Amine oxidase, copper containing 1 SEQID NOS: 875-880

AOC2 Amine oxidase, copper containing 2 (retina-specific) SEQID NOS: 881-882

AOC3 Amine oxidase, copper containing 3 SEQID NOS: 883-889

AP000721.4 SEQID NO: 890

APBB1 Amyloid beta (A4) precursor protein-binding, family SEQID NOS: 891-907

B, member 1 (Fe65)

APCDD1 Adenomatosis polyposis coli down-regulated 1 SEQID NOS: 908-913

APCS Amyloid P component, serum SEQID NO: 914

APE LA Apelin receptor early endogenous ligand SEQID NOS: 915-917

APLN Apelin SEQID NO: 918

APLP2 Amyloid beta (A4) precursor-like protein 2 SEQID NOS: 919-928

APOA1 Apolipoprotein A-l SEQID NOS: 929-933

APOA2 Apolipoprotein A-ll SEQID NOS: 934-942

APOA4 Apolipoprotein A-IV SEQID NO: 943

APOA5 Apolipoprotein A-V SEQID NOS: 944-946

APOB Apolipoprotein B SEQID NOS: 947-948

APOC1 Apolipoprotein C-l SEQID NOS: 949-957

APOC2 Apolipoprotein C-ll SEQID NOS: 958-962

APOC3 Apolipoprotein C-lll SEQID NOS: 963-966

APOC4 Apolipoprotein C-IV SEQID NOS: 967-968

APOC4- APOC4-APOC2 readthrough (NMD candidate) SEQID NOS: 969-970 APOC2

APOD Apolipoprotein D SEQID NOS: 971-974

APOE Apolipoprotein E SEQID NOS: 975-978

APOF Apolipoprotein F SEQID NO: 979

APOH Apolipoprotein H (beta-2-glycoprotein 1) SEQID NOS: 980-983

APOL1 Apolipoprotein L, 1 SEQID NOS: 984-994

APOL3 Apolipoprotein L, 3 SEQID NOS: 995- 1009

APOM Apolipoprotein M SEQID NOS: 1010-1012

APOOL Apolipoprotein O-like SEQID NOS: 1013-1015

ARCN1 Archain 1 SEQID NOS: 1016-1020

ARFIP2 ADP-ribosylation factor interacting protein 2 SEQID NOS: 1021- 1027

ARHGAP36 Rho GTPase activating protein 36 SEQID NOS: 1028- 1033

HMHA1 Histocompatibility (minor) HA-1 SEQID NOS: 1034-1042

ARHGAP6 Rho GTPase activating protein 6 SEQID NOS: 1043-1048 ARHGEF4 Rho guanine nucleotide exchange factor (GEF) 4 SEQID NOS: 1049- 1059

ARL16 ADP-ribosylation factor-like 16 SEQID NOS: 1060- 1068

ARMC5 Armadillo repeat containing 5 SEQID NOS: 1069- 1075

ARNTL Aryl hydrocarbon receptor nuclear translocator-like SEQID NOS: 1076- 1090

ARSA Arylsulfatase A SEQID NOS: 1091 - 1096

ARSB Arylsulfatase B SEQID NOS: 1097- 1100

ARSE Arylsulfatase E (chondrodysplasia punctata 1) SEQID NOS: 1101 - 1104

ARSG Arylsulfatase G SEQID NOS: 1105 - 1108

ARSI Arylsulfatase family, member 1 SEQID NOS: 1109- 1111

ARSK Arylsulfatase family, member K SEQID NOS: 1112 - 1116

ART3 ADP-ribosyltransferase 3 SEQID NOS: 1117- 1124

ART4 ADP-ribosyltransferase 4 (Dombrock blood group) SEQID NOS: 1125 - 1128

ART5 ADP-ribosyltransferase 5 SEQID NOS: 1129- 1133

ARTN Artemin SEQID NOS: 1134- 1144

ASAH1 N-acylsphingosine amidohydrolase (acid ceramidase) SEQID NOS: 1145 - 1195

1

ASAH2 N-acylsphingosine amidohydrolase (non-lysosomal SEQID NOS: 1196- 1201 ceramidase) 2

ASCL1 Achaete-scute family bHLH transcription factor 1 SEQID NO: 1202

ASIP Agouti signaling protein SEQID NOS: 1203 - 1204

ASPN Asporin SEQID NOS: 1205 - 1206

ASTL Astacin-like metallo-endopeptidase (M12 family) SEQID NO: 1207

ATAD5 ATPase family, AAA domain containing 5 SEQID NOS: 1208- 1209

AT ATI Alpha tubulin acetyltransferase 1 SEQID NOS: 1210- 1215

ATG2A Autophagy related 2A SEQID NOS: 1216- 1218

ATG5 Autophagy related 5 SEQID NOS: 1219- 1227

ATM IN ATM interactor SEQID NOS: 1228- 1231

ATP13A1 ATPase type 13A1 SEQID NOS: 1232 - 1234

ATP5F1 ATP synthase, H+ transporting, mitochondrial Fo SEQID NOS: 1235 - 1236 complex, subunit Bl

ATP6AP1 ATPase, H+ transporting, lysosomal accessory SEQID NOS: 1237- 1244 protein 1

ATP6AP2 ATPase, H+ transporting, lysosomal accessory SEQID NOS: 1245 - 1267 protein 2

ATPAF1 ATP synthase mitochondrial Fl complex assembly SEQID NOS: 1268- 1278 factor 1

AUH AU RNA binding protein/enoyl-CoA hydratase SEQID NOS: 1279- 1280

AVP Arginine vasopressin SEQID NO: 1281

AXIN2 Axin 2 SEQID NOS: 1282 - 1289

AZGP1 Alpha-2-glycoprotein 1, zinc-binding SEQID NOS: 1290- 1292

AZU1 Azurocidin 1 SEQID NOS: 1293 - 1294

B2M Beta-2-microglobulin SEQID NOS: 1295 - 1301

B3GALNT1 Beta-l,3-N-acetylgalactosaminyltransferase 1 SEQID NOS: 1302 - 1314

(globoside blood group)

B3GALNT2 Beta-l,3-N-acetylgalactosaminyltransferase 2 SEQID NOS: 1315 - 1317

B3GALT1 UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase, SEQID NO: 1318

polypeptide 1 B3GALT4 UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase, SEQID NO: 1319 polypeptide 4

B3GALT5 UDP-Gal:betaGlcNAc beta 1,3-galactosyltransferase, SEQID NOS: 1320- 1324 polypeptide 5

B3GALT6 UDP-Gal:betaGal beta 1,3-galactosyltransferase SEQID NO: 1325

polypeptide 6

B3GAT3 Beta-l,3-glucuronyltransferase 3 SEQID NOS: 1326- 1330

B3GLCT Beta 3-glucosyltransferase SEQID NO: 1331

B3GNT3 UDP-GlcNAc:betaGal beta-l,3-N- SEQID NOS: 1332 - 1335 acetylglucosaminyltransferase 3

B3GNT4 UDP-GlcNAc:betaGal beta-l,3-N- SEQID NOS: 1336- 1339 acetylglucosaminyltransferase 4

B3GNT6 UDP-GlcNAc:betaGal beta-l,3-N- SEQID NOS: 1340- 1341 acetylglucosaminyltransferase 6

B3GNT7 UDP-GlcNAc:betaGal beta-l,3-N- SEQID NO: 1342

acetylglucosaminyltransferase 7

B3GNT8 UDP-GlcNAc:betaGal beta-l,3-N- SEQID NO: 1343

acetylglucosaminyltransferase 8

B3GNT9 UDP-GlcNAc:betaGal beta-l,3-N- SEQID NO: 1344

acetylglucosaminyltransferase 9

B4GALNT1 Beta-l,4-N-acetyl-galactosaminyl transferase 1 SEQID NOS: 1345- 1356

B4GALNT3 Beta-l,4-N-acetyl-galactosaminyl transferase 3 SEQID NOS: 1357- 1358

B4GALNT4 Beta-l,4-N-acetyl-galactosaminyl transferase 4 SEQID NOS: 1359- 1361

B4GALT4 UDP-Gal:betaGlcNAc beta 1,4- galactosyltransferase, SEQID NOS: 1362 - 1375 polypeptide 4

B4GALT5 UDP-Gal:betaGlcNAc beta 1,4- galactosyltransferase, SEQID NO: 1376

polypeptide 5

B4GALT6 UDP-Gal:betaGlcNAc beta 1,4- galactosyltransferase, SEQID NOS: 1377- 1380 polypeptide 6

B4GAT1 Beta-l,4-glucuronyltransferase 1 SEQID NO: 1381

B9D1 B9 protein domain 1 SEQID NOS: 1382 - 1398

BACE2 Beta-site APP-cleaving enzyme 2 SEQID NOS: 1399- 1401

BAGE5 B melanoma antigen family, member 5 SEQID NO: 1402

BCAM Basal cell adhesion molecule (Lutheran blood group) SEQID NOS: 1403-1406

BCAN Brevican SEQID NOS: 1407-1413

BCAP29 B-cell receptor-associated protein 29 SEQID NOS: 1414-1426

BCAR1 Breast cancer anti-estrogen resistance 1 SEQID NOS: 1427-1444

BCHE Butyrylcholinesterase SEQID NOS: 1445-1449

BCKDHB Branched chain keto acid dehydrogenase El, beta SEQID NOS: 1450- 1452 polypeptide

BDNF Brain-derived neurotrophic factor SEQID NOS: 1453- 1470

BGLAP Bone gamma-carboxyglutamate (gla) protein SEQID NO: 1471

BGN Biglycan SEQID NOS: 1472-1473

BLVRB Biliverdin reductase B SEQID NOS: 1474-1478

BMP1 Bone morphogenetic protein 1 SEQID NOS: 1479-1490

BMP10 Bone morphogenetic protein 10 SEQID NO: 1491

BMP15 Bone morphogenetic protein 15 SEQID NO: 1492

BMP2 Bone morphogenetic protein 2 SEQID NO: 1493 BMP3 Bone morphogenetic protein 3 SEQID NO: 1494

BMP4 Bone morphogenetic protein 4 SEQIDNOS: 1495- 1502

BMP6 Bone morphogenetic protein 6 SEQID NO: 1503

BMP7 Bone morphogenetic protein 7 SEQID NOS: 1504- 1507

BMP8A Bone morphogenetic protein 8a SEQID NO: 1508

BMP8B Bone morphogenetic protein 8b SEQID NO: 1509

BMPER BMP binding endothelial regulator SEQID NOS: 1510-1513

BNC1 Basonuclin 1 SEQID NOS: 1514-1515

BOC BOC cell adhesion associated, oncogene regulated SEQID NOS: 1516-1526

BOD1 Biorientation of chromosomes in cell division 1 SEQID NOS: 1527- 1531

BO LAI BolA family member 1 SEQIDNOS: 1532 - 1534

BPI Bactericidal/permeability-increasing protein SEQIDNOS: 1535 - 1538

BPIFA1 BPI fold containing family A, member 1 SEQID NOS: 1539- 1542

BPIFA2 BPI fold containing family A, member 2 SEQIDNOS: 1543- 1544

BPIFA3 BPI fold containing family A, member 3 SEQIDNOS: 1545- 1546

BPIFB1 BPI fold containing family B, member 1 SEQID NOS: 1547- 1548

BPIFB2 BPI fold containing family B, member 2 SEQID NO: 1549

BPIFB3 BPI fold containing family B, member 3 SEQID NO: 1550

BPIFB4 BPI fold containing family B, member 4 SEQIDNOS: 1551 - 1552

BPIFB6 BPI fold containing family B, member 6 SEQIDNOS: 1553 - 1554

BPIFC BPI fold containing family C SEQIDNOS: 1555 - 1558

BRF1 BRF1, RNA polymerase III transcription initiation SEQID NOS: 1559- 1574 factor 90 kDa subunit

BRINP1 Bone morphogenetic protein/retinoic acid inducible SEQIDNOS: 1575 - 1576 neural-specific 1

BRINP2 Bone morphogenetic protein/retinoic acid inducible SEQID NO: 1577

neural-specific 2

BRINP3 Bone morphogenetic protein/retinoic acid inducible SEQID NOS: 1578- 1580 neural-specific 3

BSG Basigin (Ok blood group) SEQIDNOS: 1581- 1591

BSPH1 Binder of sperm protein homolog 1 SEQID NO: 1592

BST1 Bone marrow stromal cell antigen 1 SEQIDNOS: 1593 - 1597

BTBD17 BTB (POZ) domain containing 17 SEQID NO: 1598

BTD Biotinidase SEQID NOS: 1599- 1608

BTN2A2 Butyrophilin, subfamily 2, member A2 SEQID NOS: 1609- 1622

BTN3A1 Butyrophilin, subfamily 3, member Al SEQIDNOS: 1623 - 1629

BTN3A2 Butyrophilin, subfamily 3, member A2 SEQID NOS: 1630- 1640

BTN3A3 Butyrophilin, subfamily 3, member A3 SEQIDNOS: 1641-1649

RP4- Complement factor H-related protein 2 SEQID NO: 1650

608O15.3

C10orf99 Chromosome 10 open reading frame 99 SEQID NO: 1651

Cllorfl Chromosome 11 open reading frame 1 SEQIDNOS: 1652 - 1656

Cllorf24 Chromosome 11 open reading frame 24 SEQID NOS: 1657- 1659

Cllorf45 Chromosome 11 open reading frame 45 SEQID NOS: 1660- 1661

Cllorf94 Chromosome 11 open reading frame 94 SEQID NO: 1662

C12orfl0 Chromosome 12 open reading frame 10 SEQIDNOS: 1663- 1666

C12orf49 Chromosome 12 open reading frame 49 SEQID NOS: 1667- 1670 C12orf73 Chromosome 12 open reading frame 73 SEQIDNOS: 1671- 1680

C12orf76 Chromosome 12 open reading frame 76 SEQIDNOS: 1681- 1688

C14orf93 Chromosome 14 open reading frame 93 SEQID NOS: 1689- 1704

C16orf89 Chromosome 16 open reading frame 89 SEQIDNOS: 1705- 1707

C16orf90 Chromosome 16 open reading frame 90 SEQID NOS: 1708- 1709

C17orf67 Chromosome 17 open reading frame 67 SEQID NO: 1710

C17orf75 Chromosome 17 open reading frame 75 SEQIDNOS: 1711-1719

C17orf99 Chromosome 17 open reading frame 99 SEQID NOS: 1720- 1722

C18orf54 Chromosome 18 open reading frame 54 SEQIDNOS: 1723 - 1727

C19orf47 Chromosome 19 open reading frame 47 SEQID NOS: 1728- 1735

C19orf70 Chromosome 19 open reading frame 70 SEQID NOS: 1736- 1739

C1GALT1 Core 1 synthase, glycoprotein-N- SEQID NOS: 1740-1744 acetylgalactosamine 3-beta-galactosyltransferase 1

Clorfl27 Chromosome 1 open reading frame 127 SEQIDNOS: 1745-1748

Clorfl59 Chromosome 1 open reading frame 159 SEQID NOS: 1749- 1761

Clorfl98 Chromosome 1 open reading frame 198 SEQIDNOS: 1762- 1766

Clorf54 Chromosome 1 open reading frame 54 SEQID NOS: 1767- 1769

Clorf56 Chromosome 1 open reading frame 56 SEQID NO: 1770

C1QA Complement component 1, q subcomponent, A SEQID NOS: 1771 - 1773 chain

C1QB Complement component 1, q subcomponent, B SEQID NOS: 1774- 1777 chain

C1QC Complement component 1, q subcomponent, C SEQID NOS: 1778- 1780 chain

C1QL1 Complement component 1, q subcomponent-like 1 SEQID NO: 1781

C1QL2 Complement component 1, q subcomponent-like 2 SEQID NO: 1782

C1QL3 Complement component 1, q subcomponent-like 3 SEQIDNOS: 1783- 1784

C1QL4 Complement component 1, q subcomponent-like 4 SEQID NO: 1785

C1QTNF1 Clq and tumor necrosis factor related protein 1 SEQID NOS: 1786- 1795

FAM132A Family with sequence similarity 132, member A SEQID NO: 1796

C1QTNF2 Clq and tumor necrosis factor related protein 2 SEQID NO: 1797

C1QTNF3 Clq and tumor necrosis factor related protein 3 SEQID NOS: 1798- 1799

C1QTNF4 Clq and tumor necrosis factor related protein 4 SEQID NOS: 1800- 1801

C1QTNF5 Clq and tumor necrosis factor related protein 5 SEQIDNOS: 1802- 1804

C1QTNF7 Clq and tumor necrosis factor related protein 7 SEQIDNOS: 1805- 1809

C1QTNF8 Clq and tumor necrosis factor related protein 8 SEQID NOS: 1810-1811

C1QTNF9 Clq and tumor necrosis factor related protein 9 SEQIDNOS: 1812-1813

C1QTNF9B Clq and tumor necrosis factor related protein 9B SEQID NOS: 1814-1816

C1R Complement component 1, r subcomponent SEQID NOS: 1817-1825

C1RL Complement component 1, r subcomponent-like SEQID NOS: 1826- 1834

CIS Complement component 1, s subcomponent SEQIDNOS: 1835- 1844

C2 Complement component 2 SEQIDNOS: 1845- 1859

C21orf33 Chromosome 21 open reading frame 33 SEQID NOS: 1860- 1868

C21orf62 Chromosome 21 open reading frame 62 SEQID NOS: 1869- 1872

C22orfl5 Chromosome 22 open reading frame 15 SEQIDNOS: 1873- 1875

C22orf46 Chromosome 22 open reading frame 46 SEQID NO: 1876

C2CD2 C2 calcium-dependent domain containing 2 SEQID NOS: 1877- 1879 C2orf40 Chromosome 2 open reading frame 40 SEQID NOS: 1880- 1882

C2orf66 Chromosome 2 open reading frame 66 SEQID NO: 1883

C2orf69 Chromosome 2 open reading frame 69 SEQID NO: 1884

C2orf78 Chromosome 2 open reading frame 78 SEQID NO: 1885

C3 Complement component 3 SEQID NOS: 1886- 1890

C3orf33 Chromosome 3 open reading frame 33 SEQID NOS: 1891- 1895

C3orf58 Chromosome 3 open reading frame 58 SEQID NOS: 1896- 1899

C4A Complement component 4A (Rodgers blood group) SEQID NOS: 1900- 1901

C4B Complement component 4B (Chido blood group) SEQID NOS: 1902- 1903

C4BPA Complement component 4 binding protein, alpha SEQID NOS: 1904- 1906

C4BPB Complement component 4 binding protein, beta SEQID NOS: 1907- 1911

C4orf48 Chromosome 4 open reading frame 48 SEQID NOS: 1912-1913

C5 Complement component 5 SEQID NO: 1914

C5orf46 Chromosome 5 open reading frame 46 SEQID NOS: 1915-1916

C6 Complement component 6 SEQID NOS: 1917-1920

C6orfl20 Chromosome 6 open reading frame 120 SEQID NO: 1921

C6orfl5 Chromosome 6 open reading frame 15 SEQID NO: 1922

C6orf58 Chromosome 6 open reading frame 58 SEQID NO: 1923

C7 Complement component 7 SEQID NO: 1924

C7orf57 Chromosome 7 open reading frame 57 SEQID NOS: 1925 - 1929

C8A Complement component 8, alpha polypeptide SEQID NO: 1930

C8B Complement component 8, beta polypeptide SEQID NOS: 1931- 1933

C8G Complement component 8, gamma polypeptide SEQID NOS: 1934- 1935

C9 Complement component 9 SEQID NO: 1936

C9orf47 Chromosome 9 open reading frame 47 SEQID NOS: 1937- 1939

CA10 Carbonic anhydrase X SEQID NOS: 1940-1946

CA11 Carbonic anhydrase XI SEQID NOS: 1947-1948

CA6 Carbonic anhydrase VI SEQID NOS: 1949- 1953

CA9 Carbonic anhydrase IX SEQID NOS: 1954- 1955

CABLES1 Cdk5 and Abl enzyme substrate 1 SEQID NOS: 1956- 1961

CAB PI Calcium binding protein 1 SEQID NOS: 1962- 1965

CACNA2D1 Calcium channel, voltage-dependent, alpha 2/delta SEQID NOS: 1966- 1969 subunit 1

CACNA2D4 Calcium channel, voltage-dependent, alpha 2/delta SEQID NOS: 1970- 1983 subunit 4

CADM3 Cell adhesion molecule 3 SEQID NOS: 1984- 1986

CALCA Calcitonin-related polypeptide alpha SEQID NOS: 1987- 1991

CALCB Calcitonin-related polypeptide beta SEQID NOS: 1992- 1994

CALCR Calcitonin receptor SEQID NOS: 1995-2001

CALCRL Calcitonin receptor-like SEQID NOS: 2002-2006

FAM26D Family with sequence similarity 26, member D SEQID NOS: 2007-2011

CALR Calreticulin SEQID NOS: 2012 -2015

CALR3 Calreticulin 3 SEQID NOS: 2016-2017

CALU Calumenin SEQID NOS: 2018-2023

CAMK2D Calcium/calmodulin-dependent protein kinase II SEQID NOS: 2024-2035 delta

CAMP Cathelicidin antimicrobial peptide SEQID NO: 2036 CANX Calnexin SEQID NOS: 2037-2051

CAR Ml Coactivator-associated arginine methyltransferase 1 SEQID NOS: 2052 -2059

CARNS1 Carnosine synthase 1 SEQID NOS: 2060-2062

CARTPT CART prepropeptide SEQID NO: 2063

CASQ1 Calsequestrin 1 (fast-twitch, skeletal muscle) SEQID NOS: 2064-2065

CASQ2 Calsequestrin 2 (cardiac muscle) SEQID NO: 2066

CATSPERG Catsper channel auxiliary subunit gamma SEQID NOS: 2067-2074

CBLN1 Cerebellin 1 precursor SEQID NOS: 2075-2077

CBLN2 Cerebellin 2 precursor SEQID NOS: 2078-2081

CBLN3 Cerebellin 3 precursor SEQID NOS: 2082-2083

CBLN4 Cerebellin 4 precursor SEQID NO: 2084

CCBE1 Collagen and calcium binding EGF domains 1 SEQID NOS: 2085-2087

CCDC112 Coiled-coil domain containing 112 SEQID NOS: 2088-2091

CCDC129 Coiled-coil domain containing 129 SEQID NOS: 2092-2099

CCDC134 Coiled-coil domain containing 134 SEQID NOS: 2100-2101

CCDC149 Coiled-coil domain containing 149 SEQID NOS: 2102-2105

CCDC3 Coiled-coil domain containing 3 SEQID NOS: 2106-2107

CCDC80 Coiled-coil domain containing 80 SEQID NOS: 2108-2111

CCDC85A Coiled-coil domain containing 85A SEQID NO: 2112

CCDC88B Coiled-coil domain containing 88B SEQID NOS: 2113-2115

CCER2 Coiled-coil glutamate-rich protein 2 SEQID NOS: 2116-2117

CCK Cholecystokinin SEQID NOS: 2118-2120

CCL1 Chemokine (C-C motif) ligand 1 SEQID NO: 2121

CCL11 Chemokine (C-C motif) ligand 11 SEQID NO: 2122

CCL13 Chemokine (C-C motif) ligand 13 SEQID NOS: 2123-2124

CCL14 Chemokine (C-C motif) ligand 14 SEQID NOS: 2125-2128

CCL15 Chemokine (C-C motif) ligand 15 SEQID NOS: 2129-2130

CCL16 Chemokine (C-C motif) ligand 16 SEQID NOS: 2131-2133

CCL17 Chemokine (C-C motif) ligand 17 SEQID NOS: 2134-2135

CCL18 Chemokine (C-C motif) ligand 18 (pulmonary and SEQID NO: 2136

activation-regulated)

CCL19 Chemokine (C-C motif) ligand 19 SEQID NOS: 2137-2138

CCL2 Chemokine (C-C motif) ligand 2 SEQID NOS: 2139-2140

CCL20 Chemokine (C-C motif) ligand 20 SEQID NOS: 2141-2143

CCL21 Chemokine (C-C motif) ligand 21 SEQID NOS: 2144-2145

CCL22 Chemokine (C-C motif) ligand 22 SEQID NO: 2146

CCL23 Chemokine (C-C motif) ligand 23 SEQID NOS: 2147-2149

CCL24 Chemokine (C-C motif) ligand 24 SEQID NOS: 2150-2151

CCL25 Chemokine (C-C motif) ligand 25 SEQID NOS: 2152-2155

CCL26 Chemokine (C-C motif) ligand 26 SEQID NOS: 2156-2157

CCL27 Chemokine (C-C motif) ligand 27 SEQID NO: 2158

CCL28 Chemokine (C-C motif) ligand 28 SEQID NOS: 2159-2161

CCL3 Chemokine (C-C motif) ligand 3 SEQID NO: 2162

CCL3L3 Chemokine (C-C motif) ligand 3-like 3 SEQID NO: 2163

CCL4 Chemokine (C-C motif) ligand 4 SEQID NOS: 2164-2165

CCL4L2 Chemokine (C-C motif) ligand 4-like 2 SEQID NOS: 2166-2175 CCL5 Chemokine (C-C motif) ligand 5 SEQID NOS: 2176-2178

CCL7 Chemokine (C-C motif) ligand 7 SEQID NOS: 2179-2181

CCL8 Chemokine (C-C motif) ligand 8 SEQID NO: 2182

CCNB1IP1 Cyclin Bl interacting protein 1, E3 ubiquitin protein SEQID NOS: 2183-2194 ligase

CCNL1 Cyclin LI SEQID NOS: 2195 -2203

CCNL2 Cyclin L2 SEQID NOS: 2204-2211

CD14 CD14 molecule SEQID NOS: 2212 -2216

CD160 CD160 molecule SEQID NOS: 2217-2221

CD164 CD164 molecule, sialomucin SEQID NOS: 2222 -2227

CD177 CD177 molecule SEQID NOS: 2228-2230

CD1E CDle molecule SEQID NOS: 2231-2244

CD2 CD2 molecule SEQID NOS: 2245-2246

CD200 CD200 molecule SEQID NOS: 2247-2253

CD200R1 CD200 receptor 1 SEQID NOS: 2254-2258

CD22 CD22 molecule SEQID NOS: 2259-2276

CD226 CD226 molecule SEQID NOS: 2277-2284

CD24 CD24 molecule SEQID NOS: 2285 -2291

CD276 CD276 molecule SEQID NOS: 2292 -2307

CD300A CD300a molecule SEQID NOS: 2308-2312

CD300LB CD300 molecule-like family member b SEQID NOS: 2313 -2314

CD300LF CD300 molecule-like family member f SEQID NOS: 2315 -2323

CD300LG CD300 molecule-like family member g SEQID NOS: 2324-2329

CD3D CD3d molecule, delta (CD3-TCR complex) SEQID NOS: 2330-2333

CD4 CD4 molecule SEQID NOS: 2334-2336

CD40 CD40 molecule, TNF receptor superfamily member 5 SEQID NOS: 2337-2340

CD44 CD44 molecule (Indian blood group) SEQID NOS: 2341-2367

CD48 CD48 molecule SEQID NOS: 2368-2370

CD5 CD5 molecule SEQID NOS: 2371 -2372

CD55 CD55 molecule, decay accelerating factor for SEQID NOS: 2373 -2383 complement (Cromer blood group)

CD59 CD59 molecule, complement regulatory protein SEQID NOS: 2384-2394

CD5L CD5 molecule-like SEQID NO: 2395

CD6 CD6 molecule SEQID NOS: 2396-2403

CD68 CD68 molecule SEQID NOS: 2404-2407

CD7 CD7 molecule SEQID NOS: 2408-2413

CD79A CD79a molecule, immunoglobulin-associated alpha SEQID NOS: 2414-2416

CD80 CD80 molecule SEQID NOS: 2417-2419

CD86 CD86 molecule SEQID NOS: 2420-2426

CD8A CD8a molecule SEQID NOS: 2427-2430

CD8B CD8b molecule SEQID NOS: 2431-2436

CD99 CD99 molecule SEQID NOS: 2437-2445

CDC23 Cell division cycle 23 SEQID NOS: 2446-2450

CDC40 Cell division cycle 40 SEQID NOS: 2451-2453

CDC45 Cell division cycle 45 SEQID NOS: 2454-2460

CDCP1 CUB domain containing protein 1 SEQID NOS: 2461-2462

CDCP2 CUB domain containing protein 2 SEQID NOS: 2463-2464 CDH1 Cadherin 1, type 1 SEQID NOS: 2465-2472

CDH11 Cadherin 11, type 2, OB-cadherin (osteoblast) SEQID NOS: 2473-2482

CDH13 Cadherin 13 SEQID NOS: 2483-2492

CDH17 Cadherin 17, LI cadherin (liver-intestine) SEQID NOS: 2493-2497

CDH18 Cadherin 18, type 2 SEQID NOS: 2498-2504

CDH19 Cadherin 19, type 2 SEQID NOS: 2505 -2509

CDH23 Cadherin-related 23 SEQID NOS: 2510-2525

CDH5 Cadherin 5, type 2 (vascular endothelium) SEQID NOS: 2526-2533

CDHR1 Cadherin-related family member 1 SEQID NOS: 2534-2539

CDHR4 Cadherin-related family member 4 SEQID NOS: 2540-2544

CDHR5 Cadherin-related family member 5 SEQID NOS: 2545 -2551

CDKN2A Cyclin-dependent kinase inhibitor 2A SEQID NOS: 2552 -2562

CDNF Cerebral dopamine neurotrophic factor SEQID NOS: 2563 -2564

CDON Cell adhesion associated, oncogene regulated SEQID NOS: 2565 -2572

CDSN Corneodesmosin SEQID NO: 2573

CEACAM16 Carcinoembryonic antigen-related cell adhesion SEQID NOS: 2574-2575 molecule 16

CEACAM18 Carcinoembryonic antigen-related cell adhesion SEQID NO: 2576

molecule 18

CEACAM19 Carcinoembryonic antigen-related cell adhesion SEQID NOS: 2577-2583 molecule 19

CEACAM5 Carcinoembryonic antigen-related cell adhesion SEQID NOS: 2584-2591 molecule 5

CEACAM7 Carcinoembryonic antigen-related cell adhesion SEQID NOS: 2592 -2594 molecule 7

CEACAM8 Carcinoembryonic antigen-related cell adhesion SEQID NOS: 2595 -2596 molecule 8

CEL Carboxyl ester lipase SEQID NO: 2597

CELA2A Chymotrypsin-like elastase family, member 2A SEQID NO: 2598

CELA2B Chymotrypsin-like elastase family, member 2B SEQID NOS: 2599-2600

CELA3A Chymotrypsin-like elastase family, member 3A SEQID NOS: 2601-2603

CELA3B Chymotrypsin-like elastase family, member 3B SEQID NOS: 2604-2606

CEMIP Cell migration inducing protein, hyaluronan binding SEQID NOS: 2607-2611

CEP89 Centrosomal protein 89kDa SEQID NOS: 2612 -2617

CER1 Cerberus 1, DAN family BMP antagonist SEQID NO: 2618

CERCAM Cerebral endothelial cell adhesion molecule SEQID NOS: 2619-2626

CERS1 Ceramide synthase 1 SEQID NOS: 2627-2631

CES1 Carboxylesterase 1 SEQID NOS: 2632 -2637

CES3 Carboxylesterase 3 SEQID NOS: 2638-2642

CES4A Carboxylesterase 4A SEQID NOS: 2643-2648

CES5A Carboxylesterase 5A SEQID NOS: 2649-2656

CETP Cholesteryl ester transfer protein, plasma SEQID NOS: 2657-2659

CCDC108 Coiled-coil domain containing 108 SEQID NOS: 2660-2669

CFB Complement factor B SEQID NOS: 2670-2674

CFC1 Cripto, FRL-1, cryptic family 1 SEQID NOS: 2675-2677

CFC1B Cripto, FRL-1, cryptic family IB SEQID NOS: 2678-2680

CFD Complement factor D (adipsin) SEQID NOS: 2681-2682 CFDP1 Craniofacial development protein 1 SEQID NOS: 2683 -2686

CFH Complement factor H SEQID NOS: 2687-2689

CFHR1 Complement factor H-related 1 SEQID NOS: 2690-2691

CFHR2 Complement factor H-related 2 SEQID NOS: 2692 -2693

CFHR3 Complement factor H-related 3 SEQID NOS: 2694-2698

CFHR4 Complement factor H-related 4 SEQID NOS: 2699-2702

CFHR5 Complement factor H-related 5 SEQID NO: 2703

CFI Complement factor 1 SEQID NOS: 2704-2708

CFP Complement factor properdin SEQID NOS: 2709-2712

CGA Glycoprotein hormones, alpha polypeptide SEQID NOS: 2713 -2717

CGB1 Chorionic gonadotropin, beta polypeptide 1 SEQID NOS: 2718-2719

CGB2 Chorionic gonadotropin, beta polypeptide 2 SEQID NOS: 2720-2721

CGB Chorionic gonadotropin, beta polypeptide SEQID NO: 2722

CGB5 Chorionic gonadotropin, beta polypeptide 5 SEQID NO: 2723

CGB7 Chorionic gonadotropin, beta polypeptide 7 SEQID NOS: 2724-2726

CGB8 Chorionic gonadotropin, beta polypeptide 8 SEQID NO: 2727

CGREF1 Cell growth regulator with EF-hand domain 1 SEQID NOS: 2728-2735

CHAD Chondroadherin SEQID NOS: 2736-2738

CHADL Chondroadherin-like SEQID NOS: 2739-2741

CHEK2 Checkpoint kinase 2 SEQID NOS: 2742 -2763

CHGA Chromogranin A SEQID NOS: 2764-2766

CHGB Chromogranin B SEQID NOS: 2767-2768

CHI3L1 Chitinase 3-like 1 (cartilage glycoprotein-39) SEQID NOS: 2769-2770

CHI3L2 Chitinase 3-like 2 SEQID NOS: 2771 -2784

CHIA Chitinase, acidic SEQID NOS: 2785 -2793

CHID1 Chitinase domain containing 1 SEQID NOS: 2794-2812

CHIT1 Chitinase 1 (chitotriosidase) SEQID NOS: 2813 -2816

CHL1 Cell adhesion molecule Ll-like SEQID NOS: 2817-2825

CHN1 Chimerin 1 SEQID NOS: 2826-2836

CHPF Chondroitin polymerizing factor SEQID NOS: 2837-2839

CHPF2 Chondroitin polymerizing factor 2 SEQID NOS: 2840-2843

CHRD Chordin SEQID NOS: 2844-2849

CHRDL1 Chordin-like 1 SEQID NOS: 2850-2854

CHRDL2 Chordin-like 2 SEQID NOS: 2855 -2863

CHRNA2 Cholinergic receptor, nicotinic, alpha 2 (neuronal) SEQID NOS: 2864-2872

CHRNA5 Cholinergic receptor, nicotinic, alpha 5 (neuronal) SEQID NOS: 2873 -2876

CHRNB1 Cholinergic receptor, nicotinic, beta 1 (muscle) SEQID NOS: 2877-2882

CHRND Cholinergic receptor, nicotinic, delta (muscle) SEQID NOS: 2883 -2888

CHST1 Carbohydrate (keratan sulfate Gal-6) SEQID NO: 2889

sulfotransferase 1

CHST10 Carbohydrate sulfotransferase 10 SEQID NOS: 2890-2897

CHST11 Carbohydrate (chondroitin 4) sulfotransferase 11 SEQID NOS: 2898-2902

CHST13 Carbohydrate (chondroitin 4) sulfotransferase 13 SEQID NOS: 2903 -2904

CHST4 Carbohydrate (N-acetylglucosamine 6-0) SEQID NOS: 2905 -2906 sulfotransferase 4

CHST5 Carbohydrate (N-acetylglucosamine 6-0) SEQID NOS: 2907-2908 sulfotransferase 5 CHST6 Carbohydrate (N-acetylglucosamine 6-0) SEQID NOS: 2909-2910 sulfotransferase 6

CHST7 Carbohydrate (N-acetylglucosamine 6-0) SEQID NO: 2911

sulfotransferase 7

CHST8 Carbohydrate (N-acetylgalactosamine 4-0) SEQID NOS: 2912 -2915 sulfotransferase 8

CHSY1 Chondroitin sulfate synthase 1 SEQID NOS: 2916-2917

CHSY3 Chondroitin sulfate synthase 3 SEQID NO: 2918

CHTF8 Chromosome transmission fidelity factor 8 SEQID NOS: 2919-2929

CILP Cartilage intermediate layer protein, nucleotide SEQID NO: 2930

pyrophosphohydrolase

CILP2 Cartilage intermediate layer protein 2 SEQID NOS: 2931 -2932

CKLF Chemokine-like factor SEQID NOS: 2933 -2938

CKMT1A Creatine kinase, mitochondrial 1A SEQID NOS: 2939-2944

CKMT1B Creatine kinase, mitochondrial IB SEQID NOS: 2945-2954

CLCA1 Chloride channel accessory 1 SEQID NOS: 2955 -2956

CLCF1 Cardiotrophin-like cytokine factor 1 SEQID NOS: 2957-2958

CLDN15 Claudin 15 SEQID NOS: 2959-2964

CLDN7 Claudin 7 SEQID NOS: 2965-2971

CLDND1 Claudin domain containing 1 SEQID NOS: 2972-2997

CLEC11A C-type lectin domain family 11, member A SEQID NOS: 2998-3000

CLEC16A C-type lectin domain family 16, member A SEQID NOS: 3001-3006

CLEC18A C-type lectin domain family 18, member A SEQID NOS: 3007-3012

CLEC18B C-type lectin domain family 18, member B SEQID NOS: 3013 -3016

CLEC18C C-type lectin domain family 18, member C SEQID NOS: 3017-3023

CLEC19A C-type lectin domain family 19, member A SEQID NOS: 3024-3027

CLEC2B C-type lectin domain family 2, member B SEQID NOS: 3028-3029

CLEC3A C-type lectin domain family 3, member A SEQID NOS: 3030-3031

CLEC3B C-type lectin domain family 3, member B SEQID NOS: 3032 -3033

CLGN Calmegin SEQID NOS: 3034-3036

CLN5 Ceroid-lipofuscinosis, neuronal 5 SEQID NOS: 3037-3048

CLPS Colipase, pancreatic SEQID NOS: 3049-3051

CLPSL1 Colipase-like 1 SEQID NOS: 3052 -3053

CLPSL2 Colipase-like 2 SEQID NOS: 3054-3055

CLPX Caseinolytic mitochondrial matrix peptidase SEQID NOS: 3056-3058 chaperone subunit

CLSTN3 Calsyntenin 3 SEQID NOS: 3059-3065

CLU Clusterin SEQID NOS: 3066-3079

CLUL1 Clusterin-like 1 (retinal) SEQID NOS: 3080-3087

CMA1 Chymase 1, mast cell SEQID NOS: 3088-3089

CMPK1 Cytidine monophosphate (UMP-CMP) kinase 1, SEQID NOS: 3090-3093 cytosolic

CNBD1 Cyclic nucleotide binding domain containing 1 SEQID NOS: 3094-3097

CNDP1 Carnosine dipeptidase 1 (metallopeptidase M20 SEQID NOS: 3098-3100 family)

RQCD1 RCDl required for cell differentiationl homolog (S. SEQID NOS: 3101-3107 pombe)

CNPY2 Canopy FGF signaling regulator 2 SEQID NOS: 3108-3112 CNPY3 Canopy FGF signaling regulator 3 SEQID NOS: 3113 -3114

CNPY4 Canopy FGF signaling regulator 4 SEQID NOS: 3115 -3117

CNTFR Ciliary neurotrophic factor receptor SEQID NOS: 3118-3121

CNTN1 Contactin 1 SEQID NOS: 3122 -3131

CNTN2 Contactin 2 (axonal) SEQID NOS: 3132 -3143

CNTN3 Contactin 3 (plasmacytoma associated) SEQID NO: 3144

CNTN4 Contactin 4 SEQID NOS: 3145 -3153

CNTN5 Contactin 5 SEQID NOS: 3154-3159

CNTNAP2 Contactin associated protein-like 2 SEQID NOS: 3160-3163

CNTNAP3 Contactin associated protein-like 3 SEQID NOS: 3164-3168

CNTNAP3B Contactin associated protein-like 3B SEQID NOS: 3169-3177

COASY CoA synthase SEQID NOS: 3178-3187

COCH Cochlin SEQID NOS: 3188-3199

COG3 Component of oligomeric golgi complex 3 SEQID NOS: 3200-3203

COL10A1 Collagen, type X, alpha 1 SEQID NOS: 3204-3207

COL11A1 Collagen, type XI, alpha 1 SEQID NOS: 3208-3218

COL11A2 Collagen, type XI, alpha 2 SEQID NOS: 3219-3223

COL12A1 Collagen, type XII, alpha 1 SEQID NOS: 3224-3231

COL14A1 Collagen, type XIV, alpha 1 SEQID NOS: 3232 -3239

COL15A1 Collagen, type XV, alpha 1 SEQID NOS: 3240-3241

COL16A1 Collagen, type XVI, alpha 1 SEQID NOS: 3242 -3246

COL18A1 Collagen, type XVIII, alpha 1 SEQID NOS: 3247-3251

COL19A1 Collagen, type XIX, alpha 1 SEQID NOS: 3252 -3254

COL1A1 Collagen, type 1, alpha 1 SEQID NOS: 3255 -3256

COL1A2 Collagen, type 1, alpha 2 SEQID NOS: 3257-3258

COL20A1 Collagen, type XX, alpha 1 SEQID NOS: 3259-3262

COL21A1 Collagen, type XXI, alpha 1 SEQID NOS: 3263 -3268

COL22A1 Collagen, type XXII, alpha 1 SEQID NOS: 3269-3271

COL24A1 Collagen, type XXIV, alpha 1 SEQID NOS: 3272 -3275

COL26A1 Collagen, type XXVI, alpha 1 SEQID NOS: 3276-3277

COL27A1 Collagen, type XXVII, alpha 1 SEQID NOS: 3278-3280

COL28A1 Collagen, type XXVIII, alpha 1 SEQID NOS: 3281 -3285

COL2A1 Collagen, type II, alpha 1 SEQID NOS: 3286-3287

COL3A1 Collagen, type III, alpha 1 SEQID NOS: 3288-3290

COL4A1 Collagen, type IV, alpha 1 SEQID NOS: 3291 -3293

COL4A2 Collagen, type IV, alpha 2 SEQID NOS: 3294-3296

COL4A3 Collagen, type IV, alpha 3 (Goodpasture antigen) SEQID NOS: 3297-3300

COL4A4 Collagen, type IV, alpha 4 SEQID NOS: 3301 -3302

COL4A5 Collagen, type IV, alpha 5 SEQID NOS: 3303 -3309

COL4A6 Collagen, type IV, alpha 6 SEQID NOS: 3310-3315

COL5A1 Collagen, type V, alpha 1 SEQID NOS: 3316-3318

COL5A2 Collagen, type V, alpha 2 SEQID NOS: 3319-3320

COL5A3 Collagen, type V, alpha 3 SEQID NO: 3321

COL6A1 Collagen, type VI, alpha 1 SEQID NOS: 3322 -3323

COL6A2 Collagen, type VI, alpha 2 SEQID NOS: 3324-3329

COL6A3 Collagen, type VI, alpha 3 SEQID NOS: 3330-3338 COL6A5 Collagen, type VI, alpha 5 SEQID NOS: 3339-3343

COL6A6 Collagen, type VI, alpha 6 SEQID NOS: 3344-3346

COL7A1 Collagen, type VII, alpha 1 SEQID NOS: 3347-3348

COL8A1 Collagen, type VIII, alpha 1 SEQID NOS: 3349-3352

COL8A2 Collagen, type VIII, alpha 2 SEQID NOS: 3353 -3355

COL9A1 Collagen, type IX, alpha 1 SEQID NOS: 3356-3359

COL9A2 Collagen, type IX, alpha 2 SEQID NOS: 3360-3363

COL9A3 Collagen, type IX, alpha 3 SEQID NOS: 3364-3365

COLEC10 Collectin sub-family member 10 (C-type lectin) SEQID NO: 3366

COLEC11 Collectin sub-family member 11 SEQID NOS: 3367-3376

COLGALT1 Collagen beta(l-0)galactosyltransferase 1 SEQID NOS: 3377-3379

COLGALT2 Collagen beta(l-0)galactosyltransferase 2 SEQID NOS: 3380-3382

COLQ Collagen-like tail subunit (single strand of SEQID NOS: 3383 -3387 homotrimer) of asymmetric acetylcholinesterase

COMP Cartilage oligomeric matrix protein SEQID NOS: 3388-3390

COPS6 COP9 signalosome subunit 6 SEQID NOS: 3391 -3394

COQ6 Coenzyme Q6 monooxygenase SEQID NOS: 3395-3402

CORT Cortistatin SEQID NO: 3403

CP Ceruloplasmin (ferroxidase) SEQID NOS: 3404-3408

CPA1 Carboxypeptidase Al (pancreatic) SEQID NOS: 3409-3413

CPA2 Carboxypeptidase A2 (pancreatic) SEQID NOS: 3414-3415

CPA3 Carboxypeptidase A3 (mast cell) SEQID NO: 3416

CPA4 Carboxypeptidase A4 SEQID NOS: 3417-3422

CPA6 Carboxypeptidase A6 SEQID NOS: 3423-3425

CPAMD8 C3 and PZP-like, alpha-2-macroglobulin domain SEQID NOS: 3426-3431 containing 8

CPB1 Carboxypeptidase Bl (tissue) SEQID NOS: 3432-3436

CPB2 Carboxypeptidase B2 (plasma) SEQID NOS: 3437-3439

CPE Carboxypeptidase E SEQID NOS: 3440-3444

CPM Carboxypeptidase M SEQID NOS: 3445-3454

CPN1 Carboxypeptidase N, polypeptide 1 SEQID NOS: 3455-3456

CPN2 Carboxypeptidase N, polypeptide 2 SEQID NOS: 3457-3458

CPO Carboxypeptidase 0 SEQID NO: 3459

CPQ Carboxypeptidase Q SEQID NOS: 3460-3465

CPVL Carboxypeptidase, vitellogenic-like SEQID NOS: 3466-3476

CPXM1 Carboxypeptidase X (M 14 family), member 1 SEQID NO: 3477

CPXM2 Carboxypeptidase X (M14 family), member 2 SEQID NOS: 3478-3479

CPZ Carboxypeptidase Z SEQID NOS: 3480-3483

CR1L Complement component (3b/4b) receptor 1-like SEQID NOS: 3484-3485

CRB2 Crumbs family member 2 SEQID NOS: 3486-3488

CREG1 Cellular repressor of ElA-stimulated genes 1 SEQID NO: 3489

CREG2 Cellular repressor of ElA-stimulated genes 2 SEQID NO: 3490

CRELD1 Cysteine-rich with EGF-like domains 1 SEQID NOS: 3491-3496

CRELD2 Cysteine-rich with EGF-like domains 2 SEQID NOS: 3497-3501

CRH Corticotropin releasing hormone SEQID NO: 3502

CRHBP Corticotropin releasing hormone binding protein SEQID NOS: 3503 -3504

CRHR1 Corticotropin releasing hormone receptor 1 SEQID NOS: 3505 -3516 CRHR2 Corticotropin releasing hormone receptor 2 SEQID NOS: 3517-3523

CRISPl Cysteine-rich secretory protein 1 SEQID NOS: 3524-3527

CRISP2 Cysteine-rich secretory protein 2 SEQID NOS: 3528-3530

CRISP3 Cysteine-rich secretory protein 3 SEQID NOS: 3531 -3534

CRISPLD2 Cysteine-rich secretory protein LCCL domain SEQID NOS: 3535 -3542 containing 2

CRLF1 Cytokine receptor-like factor 1 SEQID NOS: 3543-3544

CRP C-reactive protein, pentraxin-related SEQID NOS: 3545-3549

CRTAC1 Cartilage acidic protein 1 SEQID NOS: 3550-3554

CRTAP Cartilage associated protein SEQID NOS: 3555 -3556

CRY2 Cryptochrome circadian clock 2 SEQID NOS: 3557-3560

CSAD Cysteine sulfinic acid decarboxylase SEQID NOS: 3561 -3573

CSF1 Colony stimulating factor 1 (macrophage) SEQID NOS: 3574-3581

CSF1R Colony stimulating factor 1 receptor SEQID NOS: 3582 -3586

CSF2 Colony stimulating factor 2 (granulocyte- SEQID NO: 3587

macrophage)

CSF2RA Colony stimulating factor 2 receptor, alpha, low- SEQID NOS: 3588-3599 affinity (granulocyte-macrophage)

CSF3 Colony stimulating factor 3 (granulocyte) SEQID NOS: 3600-3606

CSGALNACT Chondroitin sulfate N- SEQID NOS: 3607-3615 1 acetylgalactosaminyltransferase 1

CSH1 Chorionic somatomammotropin hormone 1 SEQID NOS: 3616-3619

(placental lactogen)

CSH2 Chorionic somatomammotropin hormone 2 SEQID NOS: 3620-3624

CSHL1 Chorionic somatomammotropin hormone-like 1 SEQID NOS: 3625 -3631

CSN1S1 Casein alpha si SEQID NOS: 3632 -3637

CSN2 Casein beta SEQID NO: 3638

CSN3 Casein kappa SEQID NO: 3639

CST1 Cystatin SN SEQID NOS: 3640-3641

CST11 Cystatin 11 SEQID NOS: 3642-3643

CST2 Cystatin SA SEQID NO: 3644

CST3 Cystatin C SEQID NOS: 3645-3647

CST4 Cystatin S SEQID NO: 3648

CST5 Cystatin D SEQID NO: 3649

CST6 Cystatin E/M SEQID NO: 3650

CST7 Cystatin F (leukocystatin) SEQID NO: 3651

CST8 Cystatin 8 (cystatin-related epididymal specific) SEQID NOS: 3652 -3653

CST9 Cystatin 9 (testatin) SEQID NO: 3654

CST9L Cystatin 9-like SEQID NO: 3655

CSTL1 Cystatin-like 1 SEQID NOS: 3656-3658

CT55 Cancer/testis antigen 55 SEQID NOS: 3659-3660

CTBS Chitobiase, di-N-acetyl- SEQID NOS: 3661-3663

CTGF Connective tissue growth factor SEQID NO: 3664

CTHRC1 Collagen triple helix repeat containing 1 SEQID NOS: 3665-3668

CTLA4 Cytotoxic T-lymphocyte-associated protein 4 SEQID NOS: 3669-3672

CTNS Cystinosin, lysosomal cystine transporter SEQID NOS: 3673-3680

CTRB1 Chymotrypsinogen Bl SEQID NOS: 3681-3683 CTRB2 Chymotrypsinogen B2 SEQID NOS: 3684-3687

CTRC Chymotrypsin C (caldecrin) SEQID NOS: 3688-3689

CTRL Chymotrypsin-like SEQID NOS: 3690-3692

CTSA Cathepsin A SEQID NOS: 3693-3701

CTSB Cathepsin B SEQID NOS: 3702 -3726

CTSC Cathepsin C SEQID NOS: 3727-3731

CTSD Cathepsin D SEQID NOS: 3732-3742

CTSE Cathepsin E SEQID NOS: 3743-3744

CTSF Cathepsin F SEQID NOS: 3745-3748

CTSG Cathepsin G SEQID NO: 3749

CTSH Cathepsin H SEQID NOS: 3750-3755

CTSK Cathepsin K SEQID NOS: 3756-3757

CTSL Cathepsin L SEQID NOS: 3758-3760

CTSO Cathepsin 0 SEQID NO: 3761

CTSS Cathepsin S SEQID NOS: 3762-3766

CTSV Cathepsin V SEQID NOS: 3767-3768

CTSW Cathepsin W SEQID NOS: 3769-3771

CTSZ Cathepsin Z SEQID NO: 3772

CUBN Cubilin (intrinsic factor-cobalamin receptor) SEQID NOS: 3773-3776

CUTA CutA divalent cation tolerance homolog (E. coli) SEQID NOS: 3777-3786

CX3CL1 Chemokine (C-X3-C motif) ligand 1 SEQID NOS: 3787-3790

CXADR Coxsackie virus and adenovirus receptor SEQID NOS: 3791-3795

CXCL1 Chemokine (C-X-C motif) ligand 1 (melanoma growth SEQID NO: 3796

stimulating activity, alpha)

CXCL10 Chemokine (C-X-C motif) ligand 10 SEQID NO: 3797

CXCL11 Chemokine (C-X-C motif) ligand 11 SEQID NOS: 3798-3799

CXCL12 Chemokine (C-X-C motif) ligand 12 SEQID NOS: 3800-3805

CXCL13 Chemokine (C-X-C motif) ligand 13 SEQID NO: 3806

CXCL14 Chemokine (C-X-C motif) ligand 14 SEQID NOS: 3807-3808

CXCL17 Chemokine (C-X-C motif) ligand 17 SEQID NOS: 3809-3810

CXCL2 Chemokine (C-X-C motif) ligand 2 SEQID NO: 3811

CXCL3 Chemokine (C-X-C motif) ligand 3 SEQID NO: 3812

CXCL5 Chemokine (C-X-C motif) ligand 5 SEQID NO: 3813

CXCL6 Chemokine (C-X-C motif) ligand 6 SEQID NOS: 3814-3815

CXCL8 Chemokine (C-X-C motif) ligand 8 SEQID NOS: 3816-3817

CXCL9 Chemokine (C-X-C motif) ligand 9 SEQID NO: 3818

CXorf36 Chromosome X open reading frame 36 SEQID NOS: 3819-3820

CYB5D2 Cytochrome b5 domain containing 2 SEQID NOS: 3821 -3824

CYHR1 Cysteine/histidine-rich 1 SEQID NOS: 3825 -3832

CYP17A1 Cytochrome P450, family 17, subfamily A, SEQID NOS: 3833 -3837 polypeptide 1

CYP20A1 Cytochrome P450, family 20, subfamily A, SEQID NOS: 3838-3844 polypeptide 1

CYP21A2 Cytochrome P450, family 21, subfamily A, SEQID NOS: 3845-3852 polypeptide 2

CYP26B1 Cytochrome P450, family 26, subfamily B, SEQID NOS: 3853 -3857 polypeptide 1 CYP2A6 Cytochrome P450, family 2, subfamily A, polypeptide SEQID NOS: 3858-3859 6

CYP2A7 Cytochrome P450, family 2, subfamily A, polypeptide SEQID NOS: 3860-3862

7

CYP2B6 Cytochrome P450, family 2, subfamily B, polypeptide SEQID NOS: 3863-3866

6

CYP2C18 Cytochrome P450, family 2, subfamily C, polypeptide SEQID NOS: 3867-3868

18

CYP2C19 Cytochrome P450, family 2, subfamily C, polypeptide SEQID NOS: 3869-3870

19

CYP2C8 Cytochrome P450, family 2, subfamily C, polypeptide SEQID NOS: 3871-3878

8

CYP2C9 Cytochrome P450, family 2, subfamily C, polypeptide SEQID NOS: 3879-3881

9

CYP2E1 Cytochrome P450, family 2, subfamily E, polypeptide SEQID NOS: 3882-3887

1

CYP2F1 Cytochrome P450, family 2, subfamily F, polypeptide SEQID NOS: 3888-3891

1

CYP2J2 Cytochrome P450, family 2, subfamily J, polypeptide SEQID NO: 3892

2

CYP2R1 Cytochrome P450, family 2, subfamily R, polypeptide SEQID NOS: 3893-3898

1

CYP2S1 Cytochrome P450, family 2, subfamily S, polypeptide SEQID NOS: 3899-3904

1

CYP2W1 Cytochrome P450, family 2, subfamily W, SEQID NOS: 3905-3907 polypeptide 1

CYP46A1 Cytochrome P450, family 46, subfamily A, SEQID NOS: 3908-3912 polypeptide 1

CYP4F11 Cytochrome P450, family 4, subfamily F, polypeptide SEQID NOS: 3913 -3917

11

CYP4F2 Cytochrome P450, family 4, subfamily F, polypeptide SEQID NOS: 3918-3922

2

CYR61 Cysteine-rich, angiogenic inducer, 61 SEQID NO: 3923

CYTLl Cytokine-like 1 SEQID NOS: 3924-3926

D2HGDH D-2-hydroxyglutarate dehydrogenase SEQID NOS: 3927-3935

DAG1 Dystroglycan 1 (dystrophin-associated glycoprotein SEQID NOS: 3936-3950

1)

DAND5 DAN domain family member 5, BMP antagonist SEQID NOS: 3951 -3952

DAO D-amino-acid oxidase SEQID NOS: 3953 -3958

DAZAP2 DAZ associated protein 2 SEQID NOS: 3959-3967

DBH Dopamine beta-hydroxylase (dopamine beta- SEQID NOS: 3968-3969 monooxygenase)

DBNL Drebrin-like SEQID NOS: 3970-3987

DCD Dermcidin SEQID NOS: 3988-3990

DCN Decorin SEQID NOS: 3991-4009

DDIAS DNA damage-induced apoptosis suppressor SEQID NOS: 4010-4019

DDOST Dolichyl-diphosphooligosaccharide— protein SEQID NOS: 4020-4023 glycosyltransferase subunit (non-catalytic)

DDR1 Discoidin domain receptor tyrosine kinase 1 SEQID NOS: 4024-4069 DDR2 Discoidin domain receptor tyrosine kinase 2 SEQID NOS: 4070-4075

DDT D-dopachrome tautomerase SEQID NOS: 4076-4081

DDX17 DEAD (Asp-Glu-Ala-Asp) box helicase 17 SEQID NOS: 4082-4086

DDX20 DEAD (Asp-Glu-Ala-Asp) box polypeptide 20 SEQID NOS: 4087-4089

DDX25 DEAD (Asp-Glu-Ala-Asp) box helicase 25 SEQID NOS: 4090-4096

DDX28 DEAD (Asp-Glu-Ala-Asp) box polypeptide 28 SEQID NO: 4097

DEAF1 DEAF1 transcription factor SEQID NOS: 4098-4100

DEF8 Differentially expressed in FDCP 8 homolog (mouse) SEQID NOS: 4101-4120

DEFA1 Defensin, alpha 1 SEQID NOS: 4121-4122

DEFA1B Defensin, alpha IB SEQID NO: 4123

DEFA3 Defensin, alpha 3, neutrophil-specific SEQID NO: 4124

DEFA4 Defensin, alpha 4, corticostatin SEQID NO: 4125

DEFA5 Defensin, alpha 5, Paneth cell-specific SEQID NO: 4126

DEFA6 Defensin, alpha 6, Paneth cell-specific SEQID NO: 4127

DEFB1 Defensin, beta 1 SEQID NO: 4128

DEFB103A Defensin, beta 103A SEQID NO: 4129

DEFB103B Defensin, beta 103B SEQID NO: 4130

DEFB104A Defensin, beta 104A SEQID NO: 4131

DEFB104B Defensin, beta 104B SEQID NO: 4132

DEFB105A Defensin, beta 105A SEQID NO: 4133

DEFB105B Defensin, beta 105B SEQID NO: 4134

DEFB106A Defensin, beta 106A SEQID NO: 4135

DEFB106B Defensin, beta 106B SEQID NO: 4136

DEFB107A Defensin, beta 107A SEQID NO: 4137

DEFB107B Defensin, beta 107B SEQID NO: 4138

DEFB108B Defensin, beta 108B SEQID NO: 4139

DEFB110 Defensin, beta 110 SEQID NOS: 4140-4141

DEFB113 Defensin, beta 113 SEQID NO: 4142

DEFB114 Defensin, beta 114 SEQID NO: 4143

DEFB115 Defensin, beta 115 SEQID NO: 4144

DEFB116 Defensin, beta 116 SEQID NO: 4145

DEFB118 Defensin, beta 118 SEQID NO: 4146

DEFB119 Defensin, beta 119 SEQID NOS: 4147-4149

DEFB121 Defensin, beta 121 SEQID NO: 4150

DEFB123 Defensin, beta 123 SEQID NO: 4151

DEFB124 Defensin, beta 124 SEQID NO: 4152

DEFB125 Defensin, beta 125 SEQID NO: 4153

DEFB126 Defensin, beta 126 SEQID NO: 4154

DEFB127 Defensin, beta 127 SEQID NO: 4155

DEFB128 Defensin, beta 128 SEQID NO: 4156

DEFB129 Defensin, beta 129 SEQID NO: 4157

DEFB130 Defensin, beta 130 SEQID NO: 4158

RP11- SEQID NO: 4159 1236K1.1

DEFB131 Defensin, beta 131 SEQID NO: 4160

CTD- SEQID NO: 4161

2313N18.7 DEFB132 Defensin, beta 132 SEQID NO: 4162

DEFB133 Defensin, beta 133 SEQID NO: 4163

DEFB134 Defensin, beta 134 SEQID NOS: 4164-4165

DEFB135 Defensin, beta 135 SEQID NO: 4166

DEFB136 Defensin, beta 136 SEQID NO: 4167

DEFB4A Defensin, beta 4A SEQID NO: 4168

DEFB4B Defensin, beta 4B SEQID NO: 4169

ClOorflO Chromosome 10 open reading frame 10 SEQID NOS: 4170-4171

DGCR2 DiGeorge syndrome critical region gene 2 SEQID NOS: 4172 -4175

DHH Desert hedgehog SEQID NO: 4176

DHRS4 Dehydrogenase/reductase (SDR family) member 4 SEQID NOS: 4177-4184

DHRS4L2 Dehydrogenase/reductase (SDR family) member 4 SEQID NOS: 4185 -4194 like 2

DHRS7 Dehydrogenase/reductase (SDR family) member 7 SEQID NOS: 4195 -4202

DHRS7C Dehydrogenase/reductase (SDR family) member 7C SEQID NOS: 4203 -4205

DHRS9 Dehydrogenase/reductase (SDR family) member 9 SEQID NOS: 4206-4213

DHRSX Dehydrogenase/reductase (SDR family) X-linked SEQID NOS: 4214-4218

DHX29 DEAH (Asp-Glu-Ala-His) box polypeptide 29 SEQID NOS: 4219-4221

DHX30 DEAH (Asp-Glu-Ala-His) box helicase 30 SEQID NOS: 4222 -4229

DHX8 DEAH (Asp-Glu-Ala-His) box polypeptide 8 SEQID NOS: 4230-4234

DI02 Deiodinase, iodothyronine, type II SEQID NOS: 4235 -4244

DIXDCl DIX domain containing 1 SEQID NOS: 4245 -4248

DKK1 Dickkopf WNT signaling pathway inhibitor 1 SEQID NO: 4249

DKK2 Dickkopf WNT signaling pathway inhibitor 2 SEQID NOS: 4250-4252

DKK3 Dickkopf WNT signaling pathway inhibitor 3 SEQID NOS: 4253 -4258

DKK4 Dickkopf WNT signaling pathway inhibitor 4 SEQID NO: 4259

DKKL1 Dickkopf-like 1 SEQID NOS: 4260-4265

DLG4 Discs, large homolog 4 (Drosophila) SEQID NOS: 4266-4274

DLK1 Delta-like 1 homolog (Drosophila) SEQID NOS: 4275 -4278

DLL1 Delta-like 1 (Drosophila) SEQID NOS: 4279-4280

DLL3 Delta-like 3 (Drosophila) SEQID NOS: 4281 -4283

DMBT1 Deleted in malignant brain tumors 1 SEQID NOS: 4284-4290

DMKN Dermokine SEQID NOS: 4291 -4337

DMP1 Dentin matrix acidic phosphoprotein 1 SEQID NOS: 4338-4339

DMRTA2 DMRT-like family A2 SEQID NOS: 4340-4341

DNAAF5 Dynein, axonemal, assembly factor 5 SEQID NOS: 4342 -4345

DNAH14 Dynein, axonemal, heavy chain 14 SEQID NOS: 4346-4360

DNAJB11 DnaJ (Hsp40) homolog, subfamily B, member 11 SEQID NOS: 4361 -4362

DNAJB9 DnaJ (Hsp40) homolog, subfamily B, member 9 SEQID NO: 4363

DNAJC25- DNAJC25-GNG10 readthrough SEQID NO: 4364 GNG10

DNAJC3 DnaJ (Hsp40) homolog, subfamily C, member 3 SEQID NOS: 4365 -4366

DNASE1 Deoxyribonuclease 1 SEQID NOS: 4367-4377

DNASE1L1 Deoxyribonuclease l-like 1 SEQID NOS: 4378-4388

DNASE1L2 Deoxyribonuclease l-like 2 SEQID NOS: 4389-4394

DNASE1L3 Deoxyribonuclease l-like 3 SEQID NOS: 4395 -4400

DNASE2 Deoxyribonuclease II, lysosomal SEQID NOS: 4401 -4402 DNASE2B Deoxyribonuclease II beta SEQID NOS: 4403-4404

DPEP1 Dipeptidase 1 (renal) SEQID NOS: 4405-4409

DPEP2 Dipeptidase 2 SEQID NOS: 4410-4416

DPEP3 Dipeptidase 3 SEQID NO: 4417

DPF3 D4, zinc and double PHD fingers, family 3 SEQID NOS: 4418-4424

DPP4 Dipeptidyl-peptidase 4 SEQID NOS: 4425-4429

DPP7 Dipeptidyl-peptidase 7 SEQID NOS: 4430-4435

DPT Dermatopontin SEQID NO: 4436

DRAXIN Dorsal inhibitory axon guidance protein SEQID NO: 4437

DSE Dermatan sulfate epimerase SEQID NOS: 4438-4446

DSG2 Desmoglein 2 SEQID NOS: 4447-4448

DSPP Dentin sialophosphoprotein SEQID NOS: 4449-4450

DST Dystonin SEQID NOS: 4451-4469

DU0X1 Dual oxidase 1 SEQID NOS: 4470-4474

DYNLT3 Dynein, light chain, Tctex-type 3 SEQID NOS: 4475-4477

E2F5 E2F transcription factor 5, pl30-binding SEQID NOS: 4478-4484

EBAG9 Estrogen receptor binding site associated, antigen, 9 SEQID NOS: 4485-4493

EBI3 Epstein-Barr virus induced 3 SEQID NO: 4494

ECHDC1 Ethylmalonyl-CoA decarboxylase 1 SEQID NOS: 4495-4513

ECM1 Extracellular matrix protein 1 SEQID NOS: 4514-4516

ECM2 Extracellular matrix protein 2, female organ and SEQID NOS: 4517-4520 adipocyte specific

ECS IT ECSIT signalling integrator SEQID NOS: 4521-4532

EDDM3A Epididymal protein 3A SEQID NO: 4533

EDDM3B Epididymal protein 3B SEQID NO: 4534

EDEM2 ER degradation enhancer, mannosidase alpha-like 2 SEQID NOS: 4535-4536

EDEM3 ER degradation enhancer, mannosidase alpha-like 3 SEQID NOS: 4537-4539

EDIL3 EGF-like repeats and discoidin l-like domains 3 SEQID NOS: 4540-4541

EDN1 Endothelin 1 SEQID NO: 4542

EDN2 Endothelin 2 SEQID NO: 4543

EDN3 Endothelin 3 SEQID NOS: 4544-4549

EDNRB Endothelin receptor type B SEQID NOS: 4550-4558

EFEMP1 EGF containing fibu lin-like extracellular matrix SEQID NOS: 4559-4569 protein 1

EFEMP2 EGF containing fibu lin-like extracellular matrix SEQID NOS: 4570-4581 protein 2

EFNA1 Ephrin-Al SEQID NOS: 4582-4583

EFNA2 Ephrin-A2 SEQID NO: 4584

EFNA4 Ephrin-A4 SEQID NOS: 4585-4587

EGFL6 EGF-like-domain, multiple 6 SEQID NOS: 4588-4589

EGFL7 EGF-like-domain, multiple 7 SEQID NOS: 4590-4594

EGFL8 EGF-like-domain, multiple 8 SEQID NOS: 4595-4597

EG FLAM EGF-like, fibronectin type III and laminin G domains SEQID NOS: 4598-4606

EGFR Epidermal growth factor receptor SEQID NOS: 4607-4614

EHBP1 EH domain binding protein 1 SEQID NOS: 4615-4626

EHF Ets homologous factor SEQID NOS: 4627-4636

EHMT1 Euchromatic histone-lysine N-methyltransferase 1 SEQID NOS: 4637-4662 EHMT2 Euchromatic histone-lysine N-methyltransferase 2 SEQID NOS: 4663-4667

EIF2AK1 Eukaryotic translation initiation factor 2-alpha kinase SEQID NOS: 4668-4671

1

ELANE Elastase, neutrophil expressed SEQID NOS: 4672-4673

ELN Elastin SEQID NOS: 4674-4696

ELP2 Elongator acetyltransferase complex subunit 2 SEQID NOS: 4697-4709

ELSPBP1 Epididymal sperm binding protein 1 SEQID NOS: 4710-4715

EMC1 ER membrane protein complex subunit 1 SEQID NOS: 4716-4722

EMC10 ER membrane protein complex subunit 10 SEQID NOS: 4723-4729

EMC9 ER membrane protein complex subunit 9 SEQID NOS: 4730-4733

EMCN Endomucin SEQID NOS: 4734-4738

EMID1 EMI domain containing 1 SEQID NOS: 4739-4745

EMI UNI Elastin microfibril interfacer 1 SEQID NOS: 4746-4747

EMILIN2 Elastin microfibril interfacer 2 SEQID NO: 4748

EMILIN3 Elastin microfibril interfacer 3 SEQID NO: 4749

ENAM Enamelin SEQID NO: 4750

ENDOG Endonuclease G SEQID NO: 4751

ENDOU Endonuclease, polyU-specific SEQID NOS: 4752-4754

ENHO Energy homeostasis associated SEQID NO: 4755

EN04 Enolase family member 4 SEQID NOS: 4756-4760

ENPP6 Ectonucleotide pyrophosphatase/phosphodiesterase SEQID NOS: 4761-4762

6

ENPP7 Ectonucleotide pyrophosphatase/phosphodiesterase SEQID NOS: 4763-4764

7

ENTPD5 Ectonucleoside triphosphate diphosphohydrolase 5 SEQID NOS: 4765-4769

ENTPD8 Ectonucleoside triphosphate diphosphohydrolase 8 SEQID NOS: 4770-4773

EOGT EGF domain-specific O-linked N-acetylglucosamine SEQID NOS: 4774-4781

(GlcNAc) transferase

EPCAM Epithelial cell adhesion molecule SEQID NOS: 4782-4785

EPDR1 Ependymin related 1 SEQID NOS: 4786-4789

EPGN Epithelial mitogen SEQID NOS: 4790-4798

EPHA10 EPH receptor A10 SEQID NOS: 4799-4806

EPHA3 EPH receptor A3 SEQID NOS: 4807-4809

EPHA4 EPH receptor A4 SEQID NOS: 4810-4819

EPHA7 EPH receptor A7 SEQID NOS: 4820-4821

EPHA8 EPH receptor A8 SEQID NOS: 4822-4823

EPHB2 EPH receptor B2 SEQID NOS: 4824-4828

EPHB4 EPH receptor B4 SEQID NOS: 4829-4831

EPHX3 Epoxide hydrolase 3 SEQID NOS: 4832-4835

EPO Erythropoietin SEQID NO: 4836

EPPIN Epididymal peptidase inhibitor SEQID NOS: 4837-4839

EPPIN- EPPIN-WFDC6 readthrough SEQID NO: 4840 WFDC6

EPS15 Epidermal growth factor receptor pathway substrate SEQID NOS: 4841-4843

15

EPS8L1 EPS8-like 1 SEQID NOS: 4844-4849

EPX Eosinophil peroxidase SEQID NO: 4850 EPYC Epiphycan SEQID NOS: 4851-4852

EQ.TN Equatorin, sperm acrosome associated SEQIDNOS: 4853-4855

ERAP1 Endoplasmic reticulum aminopeptidase 1 SEQID NOS: 4856-4861

ERAP2 Endoplasmic reticulum aminopeptidase 2 SEQIDNOS: 4862-4869

ERBB3 Erb-b2 receptor tyrosine kinase 3 SEQID NOS: 4870-4883

FAM132B Family with sequence similarity 132, member B SEQID NOS: 4884-4886

ERLINl ER lipid raft associated 1 SEQID NOS: 4887-4889

ERLIN2 ER lipid raft associated 2 SEQID NOS: 4890-4898

ERN1 Endoplasmic reticulum to nucleus signaling 1 SEQID NOS: 4899-4900

ERN2 Endoplasmic reticulum to nucleus signaling 2 SEQIDNOS: 4901-4905

ER01A Endoplasmic reticulum oxidoreductase alpha SEQID NOS: 4906-4912

ER01B Endoplasmic reticulum oxidoreductase beta SEQIDNOS: 4913-4915

ERP27 Endoplasmic reticulum protein 27 SEQID NOS: 4916-4917

ERP29 Endoplasmic reticulum protein 29 SEQID NOS: 4918-4921

ERP44 Endoplasmic reticulum protein 44 SEQID NO: 4922

ERV3-1 Endogenous retrovirus group 3, member 1 SEQID NO: 4923

ESM1 Endothelial cell-specific molecule 1 SEQID NOS: 4924-4926

ESRP1 Epithelial splicing regulatory protein 1 SEQID NOS: 4927-4935

EXOG Endo/exonuclease (5'-3'), endonuclease G-like SEQID NOS: 4936-4949

EXT LI Exostosin-like glycosyltransferase 1 SEQID NO: 4950

EXTL2 Exostosin-like glycosyltransferase 2 SEQIDNOS: 4951-4955

F10 Coagulation factor X SEQID NOS: 4956-4959

Fll Coagulation factor XI SEQID NOS: 4960-4964

F12 Coagulation factor XII (Hageman factor) SEQID NO: 4965

F13B Coagulation factor XIII, B polypeptide SEQID NO: 4966

F2 Coagulation factor II (thrombin) SEQID NOS: 4967-4969

F2R Coagulation factor II (thrombin) receptor SEQID NOS: 4970-4971

F2RL3 Coagulation factor II (thrombin) receptor-like 3 SEQIDNOS: 4972-4973

F5 Coagulation factor V (proaccelerin, labile factor) SEQID NOS: 4974-4975

F7 Coagulation factor VII (serum prothrombin SEQID NOS: 4976-4979 conversion accelerator)

F8 Coagulation factor VIII, procoagulant component SEQID NOS: 4980-4985

F9 Coagulation factor IX SEQID NOS: 4986-4987

FABP6 Fatty acid binding protein 6, ileal SEQID NOS: 4988-4990

FAM107B Family with sequence similarity 107, member B SEQIDNOS: 4991-5012

FAM131A Family with sequence similarity 131, member A SEQID NOS: 5013 -5021

FAM171A1 Family with sequence similarity 171, member Al SEQID NOS: 5022 -5023

FAM171B Family with sequence similarity 171, member B SEQID NOS: 5024-5025

FAM172A Family with sequence similarity 172, member A SEQID NOS: 5026-5030

FAM177A1 Family with sequence similarity 177, member Al SEQID NOS: 5031-5040

FAM180A Family with sequence similarity 180, member A SEQID NOS: 5041-5043

FAM189A1 Family with sequence similarity 189, member Al SEQID NOS: 5044-5045

FAM198A Family with sequence similarity 198, member A SEQID NOS: 5046-5048

FAM19A1 Family with sequence similarity 19 (chemokine (C-C SEQID NOS: 5049-5051 motif)-like), member Al

FAM19A2 Family with sequence similarity 19 (chemokine (C-C SEQID NOS: 5052 -5059 motif)-like), member A2 FAM19A3 Family with sequence similarity 19 (chemokine (C-C SEQID NOS: 5060-5061 motif)-like), member A3

FAM19A4 Family with sequence similarity 19 (chemokine (C-C SEQID NOS: 5062-5064 motif)-like), member A4

FAM19A5 Family with sequence similarity 19 (chemokine (C-C SEQID NOS: 5065-5068 motif)-like), member A5

FAM20A Family with sequence similarity 20, member A SEQID NOS: 5069-5072

FAM20C Family with sequence similarity 20, member C SEQID NO: 5073

FAM213A Family with sequence similarity 213, member A SEQID NOS: 5074-5079

FAM46B Family with sequence similarity 46, member B SEQID NO: 5080

FAM57A Family with sequence similarity 57, member A SEQID NOS: 5081-5086

FAM78A Family with sequence similarity 78, member A SEQID NOS: 5087-5089

FAM96A Family with sequence similarity 96, member A SEQID NOS: 5090-5094

FAM9B Family with sequence similarity 9, member B SEQID NOS: 5095-5098

FAP Fibroblast activation protein, alpha SEQID NOS: 5099-5105

FAS Fas cell surface death receptor SEQID NOS: 5106-5115

FAT1 FAT atypical cadherin 1 SEQID NOS: 5116-5122

FBLN1 Fibulin 1 SEQID NOS: 5123-5135

FBLN2 Fibulin 2 SEQID NOS: 5136-5141

FBLN5 Fibulin 5 SEQID NOS: 5142-5147

FBLN7 Fibulin 7 SEQID NOS: 5148-5153

FBN1 Fibrillin 1 SEQID NOS: 5154-5157

FBN2 Fibrillin 2 SEQID NOS: 5158-5163

FBN3 Fibrillin 3 SEQID NOS: 5164-5168

FBXW7 F-box and WD repeat domain containing 7, E3 SEQID NOS: 5169-5179 ubiquitin protein ligase

FCAR Fc fragment of IgA receptor SEQID NOS: 5180-5189

FCGBP Fc fragment of IgG binding protein SEQID NOS: 5190-5192

FCGR1B Fc fragment of IgG, high affinity lb, receptor (CD64) SEQID NOS: 5193-5198

FCGR3A Fc fragment of IgG, low affinity Ilia, receptor (CD16a) SEQID NOS: 5199-5205

FCGRT Fc fragment of IgG, receptor, transporter, alpha SEQID NOS: 5206- 5216

FCMR Fc fragment of IgM receptor SEQID NOS: 5217-5223

FCN1 Ficolin (collagen/fibrinogen domain containing) 1 SEQID NOS: 5224-5225

FCN2 Ficolin (collagen/fibrinogen domain containing SEQID NOS: 5226-5227 lectin) 2

FCN3 Ficolin (collagen/fibrinogen domain containing) 3 SEQID NOS: 5228-5229

FCRL1 Fc receptor-like 1 SEQID NOS: 5230-5232

FCRL3 Fc receptor-like 3 SEQID NOS: 5233 -5238

FCRL5 Fc receptor-like 5 SEQID NOS: 5239-5241

FCRLA Fc receptor-like A SEQID NOS: 5242 -5253

FCRLB Fc receptor-like B SEQID NOS: 5254-5258

FDCSP Follicular dendritic cell secreted protein SEQID NO: 5259

FETUB Fetuin B SEQID NOS: 5260-5266

FGA Fibrinogen alpha chain SEQID NOS: 5267-5269

FGB Fibrinogen beta chain SEQID NOS: 5270-5272

FGF10 Fibroblast growth factor 10 SEQID NOS: 5273 -5274

FGF17 Fibroblast growth factor 17 SEQID NOS: 5275 -5276 FGF18 Fibroblast growth factor 18 SEQID NO: 5277

FGF19 Fibroblast growth factor 19 SEQID NO: 5278

FGF21 Fibroblast growth factor 21 SEQID NOS: 5279-5280

FGF22 Fibroblast growth factor 22 SEQID NOS: 5281 -5282

FGF23 Fibroblast growth factor 23 SEQID NO: 5283

FGF3 Fibroblast growth factor 3 SEQID NO: 5284

FGF4 Fibroblast growth factor 4 SEQID NO: 5285

FGF5 Fibroblast growth factor 5 SEQID NOS: 5286-5288

FGF7 Fibroblast growth factor 7 SEQID NOS: 5289-5293

FGF8 Fibroblast growth factor 8 (androgen-induced) SEQID NOS: 5294-5299

FGFBP1 Fibroblast growth factor binding protein 1 SEQID NO: 5300

FGFBP2 Fibroblast growth factor binding protein 2 SEQID NO: 5301

FGFBP3 Fibroblast growth factor binding protein 3 SEQID NO: 5302

FGFR1 Fibroblast growth factor receptor 1 SEQID NOS: 5303 -5325

FGFR2 Fibroblast growth factor receptor 2 SEQID NOS: 5326-5347

FGFR3 Fibroblast growth factor receptor 3 SEQID NOS: 5348-5355

FGFR4 Fibroblast growth factor receptor 4 SEQID NOS: 5356-5365

FGFRL1 Fibroblast growth factor receptor-like 1 SEQID NOS: 5366-5371

FGG Fibrinogen gamma chain SEQID NOS: 5372 -5377

FGL1 Fibrinogen-like 1 SEQID NOS: 5378-5384

FGL2 Fibrinogen-like 2 SEQID NOS: 5385 -5386

FHL1 Four and a half LIM domains 1 SEQID NOS: 5387-5414

FH0D3 Formin homology 2 domain containing 3 SEQID NOS: 5415-5421

FIBIN Fin bud initiation factor homolog (zebrafish) SEQID NO: 5422

FICD FIC domain containing SEQID NOS: 5423-5426

FJX1 Four jointed box 1 SEQID NO: 5427

FKBP10 FK506 binding protein 10, 65 kDa SEQID NOS: 5428-5433

FKBP11 FK506 binding protein 11, 19 kDa SEQID NOS: 5434-5440

FKBP14 FK506 binding protein 14, 22 kDa SEQID NOS: 5441-5443

FKBP2 FK506 binding protein 2, 13kDa SEQID NOS: 5444-5447

FKBP7 FK506 binding protein 7 SEQID NOS: 5448-5453

FKBP9 FK506 binding protein 9, 63 kDa SEQID NOS: 5454-5457

FLT1 Fms-related tyrosine kinase 1 SEQID NOS: 5458-5466

FLT4 Fms-related tyrosine kinase 4 SEQID NOS: 5467-5471

FM01 Flavin containing monooxygenase 1 SEQID NOS: 5472-5476

FM02 Flavin containing monooxygenase 2 (non-functional) SEQID NOS: 5477-5479

FM03 Flavin containing monooxygenase 3 SEQID NOS: 5480-5482

FM05 Flavin containing monooxygenase 5 SEQID NOS: 5483-5489

FMOD Fibromodulin SEQID NO: 5490

FN1 Fibronectin 1 SEQID NOS: 5491-5503

FNDC1 Fibronectin type III domain containing 1 SEQID NOS: 5504-5505

FNDC7 Fibronectin type III domain containing 7 SEQID NOS: 5506-5507

FOCAD Focadhesin SEQID NOS: 5508-5514

F0LR2 Folate receptor 2 (fetal) SEQID NOS: 5515 -5524

F0LR3 Folate receptor 3 (gamma) SEQID NOS: 5525 -5529

F0XRED2 FAD-dependent oxidoreductase domain containing 2 SEQID NOS: 5530-5533 FP325331.1 Uncharacterized protein UNQ6126/PRO20091 SEQID NO: 5534

CH507- SEQID NOS: 5535 -5541 9B2.3

FPGS Folylpolyglutamate synthase SEQID NOS: 5542-5548

FRAS1 Fraser extracellular matrix complex subunit 1 SEQID NOS: 5549-5554

FREM1 FRAS1 related extracellular matrix 1 SEQID NOS: 5555 -5559

FREM3 FRAS1 related extracellular matrix 3 SEQID NO: 5560

FRMPD2 FERM and PDZ domain containing 2 SEQID NOS: 5561 -5564

FRZB Frizzled-related protein SEQID NO: 5565

FSHB Follicle stimulating hormone, beta polypeptide SEQID NOS: 5566-5568

FSHR Follicle stimulating hormone receptor SEQID NOS: 5569-5572

FST Follistatin SEQID NOS: 5573 -5576

FSTL1 Follistatin-like 1 SEQID NOS: 5577-5580

FSTL3 Follistatin-like 3 (secreted glycoprotein) SEQID NOS: 5581 -5586

FSTL4 Follistatin-like 4 SEQID NOS: 5587-5589

FSTL5 Follistatin-like 5 SEQID NOS: 5590-5592

FTCDNL1 Formiminotransferase cyclodeaminase N-terminal SEQID NOS: 5593 -5596 like

FUCA1 Fucosidase, alpha-L- 1, tissue SEQID NO: 5597

FUCA2 Fucosidase, alpha-L- 2, plasma SEQID NOS: 5598-5599

FURIN Furin (paired basic amino acid cleaving enzyme) SEQID NOS: 5600-5606

FUT10 Fucosyltransferase 10 (alpha (1,3) SEQID NOS: 5607-5609 fucosyltransferase)

FUT11 Fucosyltransferase 11 (alpha (1,3) SEQID NOS: 5610-5611 fucosyltransferase)

FXN Frataxin SEQID NOS: 5612 -5619

FXR1 Fragile X mental retardation, autosomal homolog 1 SEQID NOS: 5620-5632

FXYD3 FXYD domain containing ion transport regulator 3 SEQID NOS: 5633-5645

GABBR1 Gamma-aminobutyric acid (GABA) B receptor, 1 SEQID NOS: 5646-5657

GABRA1 Gamma-aminobutyric acid (GABA) A receptor, alpha SEQID NOS: 5658-5673

1

GABRA2 Gamma-aminobutyric acid (GABA) A receptor, alpha SEQID NOS: 5674-5688

2

GABRA5 Gamma-aminobutyric acid (GABA) A receptor, alpha SEQID NOS: 5689-5697

5

GABRG3 Gamma-aminobutyric acid (GABA) A receptor, SEQID NOS: 5698-5703 gamma 3

GABRP Gamma-aminobutyric acid (GABA) A receptor, pi SEQID NOS: 5704-5712

GAL Galanin/GMAP prepropeptide SEQID NO: 5713

GAL3ST1 Galactose-3-O-sulfotransferase 1 SEQID NOS: 5714-5735

GAL3ST2 Galactose-3-O-sulfotransferase 2 SEQID NO: 5736

GAL3ST3 Galactose-3-O-sulfotransferase 3 SEQID NOS: 5737-5738

GALC Galactosylceramidase SEQID NOS: 5739-5748

GALNS Galactosamine (N-acetyl)-6-sulfatase SEQID NOS: 5749-5754

GALNT10 Polypeptide N-acetylgalactosaminyltransferase 10 SEQID NOS: 5755 -5758

GALNT12 Polypeptide N-acetylgalactosaminyltransferase 12 SEQID NOS: 5759-5760

GALNT15 Polypeptide N-acetylgalactosaminyltransferase 15 SEQID NOS: 5761-5764

GALNT2 Polypeptide N-acetylgalactosaminyltransferase 2 SEQID NO: 5765 GALNT6 Polypeptide N-acetylgalactosaminyltransferase 6 SEQID NOS: 5766-5777

GALNT8 Polypeptide N-acetylgalactosaminyltransferase 8 SEQID NOS: 5778-5781

GALNTL6 Polypeptide N-acetylgalactosaminyltransferase-like SEQID NOS: 5782-5785

6

GALP Galanin-like peptide SEQID NOS: 5786-5788

GANAB Glucosidase, alpha; neutral AB SEQID NOS: 5789-5797

GARS Glycyl-tRNA synthetase SEQID NOS: 5798-5801

GAS1 Growth arrest-specific 1 SEQID NO: 5802

GAS6 Growth arrest-specific 6 SEQID NO: 5803

GAST Gastrin SEQID NO: 5804

PDDC1 Parkinson disease 7 domain containing 1 SEQID NOS: 5805-5813

GBA Glucosidase, beta, acid SEQID NOS: 5814-5817

GBGT1 Globoside alpha-l,3-N- SEQID NOS: 5818-5826 acetylgalactosaminyltransferase 1

GC Group-specific component (vitamin D binding SEQID NOS: 5827-5831 protein)

GCG Glucagon SEQID NOS: 5832 -5833

GCGR Glucagon receptor SEQID NOS: 5834-5836

GCNT7 Glucosaminyl (N-acetyl) transferase family member SEQID NOS: 5837-5838

7

GCSH Glycine cleavage system protein H (aminomethyl SEQID NOS: 5839-5847 carrier)

GDF1 Growth differentiation factor 1 SEQID NO: 5848

GDF10 Growth differentiation factor 10 SEQID NO: 5849

GDF11 Growth differentiation factor 11 SEQID NOS: 5850-5851

GDF15 Growth differentiation factor 15 SEQID NOS: 5852 -5854

GDF2 Growth differentiation factor 2 SEQID NO: 5855

GDF3 Growth differentiation factor 3 SEQID NO: 5856

GDF5 Growth differentiation factor 5 SEQID NOS: 5857-5858

GDF6 Growth differentiation factor 6 SEQID NOS: 5859-5861

GDF7 Growth differentiation factor 7 SEQID NO: 5862

GDF9 Growth differentiation factor 9 SEQID NOS: 5863-5867

GDNF Glial cell derived neurotrophic factor SEQID NOS: 5868-5875

GF0D2 Glucose-fructose oxidoreductase domain containing SEQID NOS: 5876-5881

2

GFPT2 Glutamine-fructose-6-phosphate transaminase 2 SEQID NOS: 5882-5884

GFRA2 GDNF family receptor alpha 2 SEQID NOS: 5885-5891

GFRA4 GDNF family receptor alpha 4 SEQID NOS: 5892-5894

GGA2 Golgi-associated, gamma adaptin ear containing, SEQID NOS: 5895-5903

ARF binding protein 2

GGH Gamma-glutamyl hydrolase (conjugase, SEQID NO: 5904

folylpolygammaglutamyl hydrolase)

GGT1 Gamma-glutamyltransferase 1 SEQID NOS: 5905 -5927

GGT5 Gamma-glutamyltransferase 5 SEQID NOS: 5928-5932

GH1 Growth hormone 1 SEQID NOS: 5933 -5937

GH2 Growth hormone 2 SEQID NOS: 5938-5942

GHDC GH3 domain containing SEQID NOS: 5943-5950

GHRH Growth hormone releasing hormone SEQID NOS: 5951 -5953 GHRHR Growth hormone releasing hormone receptor SEQID NOS: 5954-5959

GHRL Ghrelin/obestatin prepropeptide SEQID NOS: 5960-5970

GIF Gastric intrinsic factor (vitamin B synthesis) SEQID NOS: 5971-5972

GIP Gastric inhibitory polypeptide SEQID NO: 5973

GKN1 Gastrokine 1 SEQID NO: 5974

GKN2 Gastrokine 2 SEQID NOS: 5975-5976

GLA Galactosidase, alpha SEQID NOS: 5977-5978

GLB1 Galactosidase, beta 1 SEQID NOS: 5979-5987

GLB1L Galactosidase, beta 1-like SEQID NOS: 5988-5995

GLB1L2 Galactosidase, beta 1-like 2 SEQID NOS: 5996-5997

GLCE Glucuronic acid epimerase SEQID NOS: 5998-5999

GLG1 Golgi glycoprotein 1 SEQID NOS: 6000-6007

GLIPRl GLI pathogenesis-related 1 SEQID NOS: 6008-6011

GLIPRILI GLI pathogenesis-related 1 like 1 SEQID NOS: 6012-6015

GLIS3 GLIS family zinc finger 3 SEQID NOS: 6016-6024

GLMP Glycosylated lysosomal membrane protein SEQID NOS: 6025-6033

GLRB Glycine receptor, beta SEQID NOS: 6034-6039

GLS Glutaminase SEQID NOS: 6040-6047

GLT6D1 Glycosyltransferase 6 domain containing 1 SEQID NOS: 6048-6049

GLTPD2 Glycolipid transfer protein domain containing 2 SEQID NO: 6050

GLUD1 Glutamate dehydrogenase 1 SEQID NO: 6051

GM2A GM2 ganglioside activator SEQID NOS: 6052-6054

GML Glycosylphosphatidylinositol anchored molecule like SEQID NOS: 6055-6056

GNAS GNAS complex locus SEQID NOS: 6057-6078

GNLY Granulysin SEQID NOS: 6079-6082

GNPTG N-acetylglucosamine-l-phosphate transferase, SEQID NOS: 6083-6087 gamma subunit

GNRH1 Gonadotropin-releasing hormone 1 (luteinizing- SEQID NOS: 6088-6089 releasing hormone)

GNRH2 Gonadotropin-releasing hormone 2 SEQID NOS: 6090-6093

GNS Glucosamine (N-acetyl)-6-sulfatase SEQID NOS: 6094-6099

G0LM1 Golgi membrane protein 1 SEQID NOS: 6100-6104

GORAB Golgin, RAB6-interacting SEQID NOS: 6105-6107

GOT2 Glutamic-oxaloacetic transaminase 2, mitochondrial SEQID NOS: 6108-6110

GP2 Glycoprotein 2 (zymogen granule membrane) SEQID NOS: 6111-6119

GP6 Glycoprotein VI (platelet) SEQID NOS: 6120-6123

GPC2 Glypican 2 SEQID NOS: 6124-6125

GPC5 Glypican 5 SEQID NOS: 6126-6128

GPC6 Glypican 6 SEQID NOS: 6129-6130

GPD2 Glycerol-3-phosphate dehydrogenase 2 SEQID NOS: 6131-6139

(mitochondrial)

GPER1 G protein-coupled estrogen receptor 1 SEQID NOS: 6140-6146

GPHA2 Glycoprotein hormone alpha 2 SEQID NOS: 6147-6149

GPHB5 Glycoprotein hormone beta 5 SEQID NOS: 6150-6151

GPIHBP1 Glycosylphosphatidylinositol anchored high density SEQID NO: 6152

lipoprotein binding protein 1 GPLD1 Glycosylphosphatidylinositol specific phospholipase SEQID NO: 6153 Dl

GPNMB Glycoprotein (transmembrane) nmb SEQID NOS: 6154-6156

GPR162 G protein-coupled receptor 162 SEQID NOS: 6157-6160

GPX3 Glutathione peroxidase 3 SEQID NOS: 6161-6168

GPX4 Glutathione peroxidase 4 SEQID NOS: 6169-6179

GPX5 Glutathione peroxidase 5 SEQID NOS: 6180-6181

GPX6 Glutathione peroxidase 6 SEQID NOS: 6182-6184

GPX7 Glutathione peroxidase 7 SEQID NO: 6185

GREM1 Gremlin 1, DAN family BMP antagonist SEQID NOS: 6186-6188

GREM2 Gremlin 2, DAN family BMP antagonist SEQID NO: 6189

GRHL3 Grainyhead-like transcription factor 3 SEQID NOS: 6190-6195

GRIA2 Glutamate receptor, ionotropic, AMPA 2 SEQID NOS: 6196-6207

GRIA3 Glutamate receptor, ionotropic, AMPA 3 SEQID NOS: 6208-6213

GRIA4 Glutamate receptor, ionotropic, AMPA 4 SEQID NOS: 6214-6225

GRIK2 Glutamate receptor, ionotropic, kainate 2 SEQID NOS: 6226-6234

GRIN2B Glutamate receptor, ionotropic, N-methyl D- SEQID NOS: 6235 -6238 aspartate 2B

GRM2 Glutamate receptor, metabotropic 2 SEQID NOS: 6239-6242

GRM3 Glutamate receptor, metabotropic 3 SEQID NOS: 6243-6247

GRM5 Glutamate receptor, metabotropic 5 SEQID NOS: 6248-6252

GRN Granulin SEQID NOS: 6253 -6268

GRP Gastrin-releasing peptide SEQID NOS: 6269-6273

DFNA5 Deafness, autosomal dominant 5 SEQID NOS: 6274-6282

GSG1 Germ cell associated 1 SEQID NOS: 6283-6291

GSN Gelsolin SEQID NOS: 6292-6300

GTDC1 Glycosyltransferase-like domain containing 1 SEQID NOS: 6301-6314

GTPBP10 GTP-binding protein 10 (putative) SEQID NOS: 6315 -6323

GUCA2A Guanylate cyclase activator 2A (guanylin) SEQID NO: 6324

GUCA2B Guanylate cyclase activator 2B (uroguanylin) SEQID NO: 6325

GUSB Glucuronidase, beta SEQID NOS: 6326-6330

GVQW1 GVQW motif containing 1 SEQID NO: 6331

GXYLT1 Glucoside xylosyltransferase 1 SEQID NOS: 6332 -6333

GXYLT2 Glucoside xylosyltransferase 2 SEQID NOS: 6334-6336

GYPB Glycophorin B (MNS blood group) SEQID NOS: 6337-6345

GZMA Granzyme A (granzyme 1, cytotoxic T-lymphocyte- SEQID NO: 6346

associated serine esterase 3)

GZMB Granzyme B (granzyme 2, cytotoxic T-lymphocyte- SEQID NOS: 6347-6355 associated serine esterase 1)

GZMH Granzyme H (cathepsin G-like 2, protein h-CCPX) SEQID NOS: 6356-6358

GZMK Granzyme K (granzyme 3; tryptase II) SEQID NO: 6359

GZMM Granzyme M (lymphocyte met-ase 1) SEQID NOS: 6360-6361

H6PD Hexose-6-phosphate dehydrogenase (glucose 1- SEQID NOS: 6362-6363 dehydrogenase)

HABP2 Hyaluronan binding protein 2 SEQID NOS: 6364-6365 HADHB Hydroxyacyl-CoA dehydrogenase/3-ketoacyl-CoA SEQID NOS: 6366-6372 thiolase/enoyl-CoA hydratase (trifunctional protein),

beta subunit

HAMP Hepcidin antimicrobial peptide SEQID NOS: 6373 -6374

HAPLN1 Hyaluronan and proteoglycan link protein 1 SEQID NOS: 6375 -6381

HAPLN2 Hyaluronan and proteoglycan link protein 2 SEQID NOS: 6382 -6383

HAPLN3 Hyaluronan and proteoglycan link protein 3 SEQID NOS: 6384-6387

HAPLN4 Hyaluronan and proteoglycan link protein 4 SEQID NO: 6388

HARS2 Histidyl-tRNA synthetase 2, mitochondrial SEQID NOS: 6389-6404

HAVCR1 Hepatitis A virus cellular receptor 1 SEQID NOS: 6405 -6409

HCCS Holocytochrome c synthase SEQID NOS: 6410-6412

HCRT Hypocretin (orexin) neuropeptide precursor SEQID NO: 6413

CECR5 Cat eye syndrome chromosome region, candidate 5 SEQID NOS: 6414-6416

HEATR5A HEAT repeat containing 5A SEQID NOS: 6417-6423

HEPH Hephaestin SEQID NOS: 6424-6431

HEXA Hexosaminidase A (alpha polypeptide) SEQID NOS: 6432 -6441

HEXB Hexosaminidase B (beta polypeptide) SEQID NOS: 6442 -6447

HFE2 Hemochromatosis type 2 (juvenile) SEQID NOS: 6448-6454

HGF Hepatocyte growth factor (hepapoietin A; scatter SEQID NOS: 6455 -6465 factor)

HGFAC HGF activator SEQID NOS: 6466-6467

HHIP Hedgehog interacting protein SEQID NOS: 6468-6469

HHIPL1 HHIP-like 1 SEQID NOS: 6470-6471

HHIPL2 HHIP-like 2 SEQID NO: 6472

HHLA1 HERV-H LTR-associating 1 SEQID NOS: 6473 -6474

HHLA2 HERV-H LTR-associating 2 SEQID NOS: 6475 -6485

HIBADH 3-hydroxyisobutyrate dehydrogenase SEQID NOS: 6486-6488

HINT2 Histidine triad nucleotide binding protein 2 SEQID NO: 6489

HLA-A Major histocompatibility complex, class 1, A SEQID NOS: 6490-6494

HLA-C Major histocompatibility complex, class 1, C SEQID NOS: 6495 -6499

H LA- DO A Major histocompatibility complex, class II, DO alpha SEQID NOS: 6500-6501

HLA-DPA1 Major histocompatibility complex, class II, DP alpha SEQID NOS: 6502 -6505

1

HLA-DQA1 Major histocompatibility complex, class II, DQ alpha SEQID NOS: 6506-6511

1

HLA-DQB1 Major histocompatibility complex, class II, DQ beta 1 SEQID NOS: 6512 -6517

HLA-DQB2 Major histocompatibility complex, class II, DQ beta 2 SEQID NOS: 6518-6521

HMCN1 Hemicentin 1 SEQID NOS: 6522 -6523

HMCN2 Hemicentin 2 SEQID NOS: 6524-6527

HMGCL 3-hydroxymethyl-3-methylglutaryl-CoA lyase SEQID NOS: 6528-6531

HMSD Histocompatibility (minor) serpin domain containing SEQID NOS: 6532 -6533

HP Haptoglobin SEQID NOS: 6534-6547

HPR Haptoglobin-related protein SEQID NOS: 6548-6550

HPSE Heparanase SEQID NOS: 6551 -6557

HPSE2 Heparanase 2 (inactive) SEQID NOS: 6558-6563

HPX Hemopexin SEQID NOS: 6564-6565

HRC Histidine rich calcium binding protein SEQID NOS: 6566-6568 HRG Histidine-rich glycoprotein SEQID NO: 6569

HS2ST1 Heparan sulfate 2-O-sulfotransferase 1 SEQID NOS: 6570-6572

HS3ST1 Heparan sulfate (glucosamine) 3-O-sulfotransferase SEQID NOS: 6573-6575

1

HS6ST1 Heparan sulfate 6-O-sulfotransferase 1 SEQID NO: 6576

HS6ST3 Heparan sulfate 6-O-sulfotransferase 3 SEQID NOS: 6577-6578

HSD11B1L Hydroxysteroid (11-beta) dehydrogenase 1-like SEQID NOS: 6579-6597

HSD17B11 Hydroxysteroid (17-beta) dehydrogenase 11 SEQID NOS: 6598-6599

HSD17B7 Hydroxysteroid (17-beta) dehydrogenase 7 SEQID NOS: 6600-6604

HSP90B1 Heat shock protein 90kDa beta (Grp94), member 1 SEQID NOS: 6605-6610

HSPA13 Heat shock protein 70kDa family, member 13 SEQID NO: 6611

HSPA5 Heat shock 70kDa protein 5 (glucose-regulated SEQID NO: 6612

protein, 78kDa)

HSPG2 Heparan sulfate proteoglycan 2 SEQID NOS: 6613-6617

HTATIP2 HIV-1 Tat interactive protein 2, 30kDa SEQID NOS: 6618-6625

HTN1 Histatin 1 SEQID NOS: 6626-6628

HTN3 Histatin 3 SEQID NOS: 6629-6631

HTRA1 HtrA serine peptidase 1 SEQID NOS: 6632-6633

HTRA3 HtrA serine peptidase 3 SEQID NOS: 6634-6635

HTRA4 HtrA serine peptidase 4 SEQID NO: 6636

HYAL1 Hyaluronoglucosaminidase 1 SEQID NOS: 6637-6645

HYAL2 Hyaluronoglucosaminidase 2 SEQID NOS: 6646-6654

HYAL3 Hyaluronoglucosaminidase 3 SEQID NOS: 6655-6661

HY0U1 Hypoxia up-regulated 1 SEQID NOS: 6662-6676

IAPP Islet amyloid polypeptide SEQID NOS: 6677-6681

IBSP Integrin-binding sialoprotein SEQID NO: 6682

ICAM1 Intercellular adhesion molecule 1 SEQID NOS: 6683-6685

ICAM2 Intercellular adhesion molecule 2 SEQID NOS: 6686-6696

ICAM4 Intercellular adhesion molecule 4 (Landsteiner- SEQID NOS: 6697-6699

Wiener blood group)

ID1 Inhibitor of DNA binding 1, dominant negative helix- SEQID NOS: 6700-6701 loop-helix protein

IDE Insulin-degrading enzyme SEQID NOS: 6702-6705

IDNK IdnK, gluconokinase homolog (E. coli) SEQID NOS: 6706-6711

IDS Iduronate 2-sulfatase SEQID NOS: 6712-6717

IDUA Iduronidase, alpha-L- SEQID NOS: 6718-6723

IFI27L2 Interferon, alpha-inducible protein 27-like 2 SEQID NOS: 6724-6725

IFI30 Interferon, gamma-inducible protein 30 SEQID NOS: 6726-6727

IFNA1 Interferon, alpha 1 SEQID NO: 6728

IFNA10 Interferon, alpha 10 SEQID NO: 6729

IFNA13 Interferon, alpha 13 SEQID NOS: 6730-6731

IFNA14 Interferon, alpha 14 SEQID NO: 6732

IFNA16 Interferon, alpha 16 SEQID NO: 6733

IFNA17 Interferon, alpha 17 SEQID NO: 6734

IFNA2 Interferon, alpha 2 SEQID NO: 6735

IFNA21 Interferon, alpha 21 SEQID NO: 6736

IFNA4 Interferon, alpha 4 SEQID NO: 6737 IFNA5 Interferon, alpha 5 SEQID NO: 6738

IFNA6 Interferon, alpha 6 SEQID NOS: 6739-6740

IFNA7 Interferon, alpha 7 SEQID NO: 6741

IFNA8 Interferon, alpha 8 SEQID NO: 6742

IFNAR1 Interferon (alpha, beta and omega) receptor 1 SEQID NOS: 6743-6744

IFNB1 Interferon, beta 1, fibroblast SEQID NO: 6745

IFNE Interferon, epsilon SEQID NO: 6746

IFNG Interferon, gamma SEQID NO: 6747

IFNGR1 Interferon gamma receptor 1 SEQID NOS: 6748-6758

IFNL1 Interferon, lambda 1 SEQID NO: 6759

IFNL2 Interferon, lambda 2 SEQID NO: 6760

IFNL3 Interferon, lambda 3 SEQID NOS: 6761-6762

IFNLR1 Interferon, lambda receptor 1 SEQID NOS: 6763-6767

IFNW1 Interferon, omega 1 SEQID NO: 6768

IGF1 Insulin-like growth factor 1 (somatomedin C) SEQID NOS: 6769-6774

IGF2 Insulin-like growth factor 2 SEQID NOS: 6775-6782

IGFALS Insulin-like growth factor binding protein, acid labile SEQID NOS: 6783-6785 subunit

IGFBP1 Insulin-like growth factor binding protein 1 SEQID NOS: 6786-6788

IGFBP2 Insulin-like growth factor binding protein 2, 36kDa SEQID NOS: 6789-6792

IGFBP3 Insulin-like growth factor binding protein 3 SEQID NOS: 6793-6800

IGFBP4 Insulin-like growth factor binding protein 4 SEQID NO: 6801

IGFBP5 Insulin-like growth factor binding protein 5 SEQID NOS: 6802-6803

IGFBP6 Insulin-like growth factor binding protein 6 SEQID NOS: 6804-6806

IGFBP7 Insulin-like growth factor binding protein 7 SEQID NOS: 6807-6808

IGFBPL1 Insulin-like growth factor binding protein-like 1 SEQID NO: 6809

IGFL1 IGF-like family member 1 SEQID NO: 6810

IGFL2 IGF-like family member 2 SEQID NOS: 6811-6813

IGFL3 IGF-like family member 3 SEQID NO: 6814

IGFLR1 IGF-like family receptor 1 SEQID NOS: 6815-6823

IGIP IgA-inducing protein SEQID NO: 6824

IGL0N5 IgLON family member 5 SEQID NO: 6825

IGSF1 Immunoglobulin superfamily, member 1 SEQID NOS: 6826-6831

IGSF10 Immunoglobulin superfamily, member 10 SEQID NOS: 6832-6833

IGSF11 Immunoglobulin superfamily, member 11 SEQID NOS: 6834-6841

IGSF21 Immunoglobin superfamily, member 21 SEQID NO: 6842

IGSF8 Immunoglobulin superfamily, member 8 SEQID NOS: 6843-6846

IGSF9 Immunoglobulin superfamily, member 9 SEQID NOS: 6847-6849

IHH Indian hedgehog SEQID NO: 6850

IL10 Interleukin 10 SEQID NOS: 6851-6852

IL11 Interleukin 11 SEQID NOS: 6853-6856

IL11RA Interleukin 11 receptor, alpha SEQID NOS: 6857-6867

IL12B Interleukin 12B SEQID NO: 6868

IL12RB1 Interleukin 12 receptor, beta 1 SEQID NOS: 6869-6874

IL12RB2 Interleukin 12 receptor, beta 2 SEQID NOS: 6875-6879

IL13 Interleukin 13 SEQID NOS: 6880-6881 IL13RA1 nterleukin 13 receptor, alpha 1 SEQID NOS: 6882 -6883

IL15RA nterleukin 15 receptor, alpha SEQID NOS: 6884-6901

IL17A nterleukin 17A SEQID NO: 6902

IL17B nterleukin 17B SEQID NO: 6903

IL17C nterleukin 17C SEQID NO: 6904

IL17D nterleukin 17D SEQID NOS: 6905 -6907

IL17F nterleukin 17F SEQID NO: 6908

IL17RA nterleukin 17 receptor A SEQID NOS: 6909-6910

IL17RC nterleukin 17 receptor C SEQID NOS: 6911 -6926

IL17RE nterleukin 17 receptor E SEQID NOS: 6927-6933

IL18BP nterleukin 18 binding protein SEQID NOS: 6934-6944

IL18R1 nterleukin 18 receptor 1 SEQID NOS: 6945 -6948

IL18RAP nterleukin 18 receptor accessory protein SEQID NOS: 6949-6951

IL19 nterleukin 19 SEQID NOS: 6952 -6954

IL1R1 nterleukin 1 receptor, type 1 SEQID NOS: 6955 -6967

IL1R2 nterleukin 1 receptor, type II SEQID NOS: 6968-6971

IL1RAP nterleukin 1 receptor accessory protein SEQID NOS: 6972 -6985

IL1RL1 nterleukin 1 receptor-like 1 SEQID NOS: 6986-6991

IL1RL2 nterleukin 1 receptor-like 2 SEQID NOS: 6992 -6994

IL1RN nterleukin 1 receptor antagonist SEQID NOS: 6995 -6999

IL2 nterleukin 2 SEQID NO: 7000

IL20 nterleukin 20 SEQID NOS: 7001 -7003

IL20RA nterleukin 20 receptor, alpha SEQID NOS: 7004-7010

IL21 nterleukin 21 SEQID NOS: 7011 -7012

IL22 nterleukin 22 SEQID NOS: 7013 -7014

IL22RA2 nterleukin 22 receptor, alpha 2 SEQID NOS: 7015 -7017

IL23A nterleukin 23, alpha subunit pl9 SEQID NO: 7018

IL24 nterleukin 24 SEQID NOS: 7019-7024

IL25 nterleukin 25 SEQID NOS: 7025 -7026

IL26 nterleukin 26 SEQID NO: 7027

IL27 nterleukin 27 SEQID NOS: 7028-7029

IL2RB nterleukin 2 receptor, beta SEQID NOS: 7030-7034

IL3 nterleukin 3 SEQID NO: 7035

IL31 nterleukin 31 SEQID NO: 7036

IL31RA nterleukin 31 receptor A SEQID NOS: 7037-7044

IL32 nterleukin 32 SEQID NOS: 7045 -7074

IL34 nterleukin 34 SEQID NOS: 7075 -7078

IL3RA nterleukin 3 receptor, alpha (low affinity) SEQID NOS: 7079-7081

IL4 nterleukin 4 SEQID NOS: 7082 -7084

IL4I1 nterleukin 4 induced 1 SEQID NOS: 7085 -7092

IL4R nterleukin 4 receptor SEQID NOS: 7093 -7106

IL5 nterleukin 5 SEQID NOS: 7107-7108

IL5RA nterleukin 5 receptor, alpha SEQID NOS: 7109-7118

IL6 nterleukin 6 SEQID NOS: 7119-7125

IL6R nterleukin 6 receptor SEQID NOS: 7126-7131

IL6ST nterleukin 6 signal transducer SEQID NOS: 7132 -7141 IL7 Interleukin 7 SEQID NOS: 7142-7149

IL7R Interleukin 7 receptor SEQID NOS: 7150-7156

IL9 Interleukin 9 SEQID NO: 7157

ILDR1 Immunoglobulin-like domain containing receptor 1 SEQID NOS: 7158-7162

ILDR2 Immunoglobulin-like domain containing receptor 2 SEQID NOS: 7163-7169

IMP4 IMP4, U3 small nucleolar ribonucleoprotein SEQID NOS: 7170-7175

IMPG1 Interphotoreceptor matrix proteoglycan 1 SEQID NOS: 7176-7179

INHA Inhibin, alpha SEQID NO: 7180

INHBA Inhibin, beta A SEQID NOS: 7181-7183

INHBB Inhibin, beta B SEQID NO: 7184

INHBC Inhibin, beta C SEQID NO: 7185

INHBE Inhibin, beta E SEQID NOS: 7186-7187

INPP5A Inositol polyphosphate-5-phosphatase A SEQID NOS: 7188-7192

INS Insulin SEQID NOS: 7193-7197

INS-IGF2 INS-IGF2 readthrough SEQID NOS: 7198-7199

INSL3 Insulin-like 3 (Leydig cell) SEQID NOS: 7200-7202

INSL4 Insulin-like 4 (placenta) SEQID NO: 7203

INSL5 Insulin-like 5 SEQID NO: 7204

INSL6 Insulin-like 6 SEQID NO: 7205

INTS3 Integrator complex subunit 3 SEQID NOS: 7206-7211

IP011 Importin 11 SEQID NOS: 7212 -7220

IP09 Importin 9 SEQID NOS: 7221 -7222

IQCF6 IQ motif containing F6 SEQID NOS: 7223 -7224

IRAK3 lnterleukin-1 receptor-associated kinase 3 SEQID NOS: 7225 -7227

IRS4 Insulin receptor substrate 4 SEQID NO: 7228

ISLR Immunoglobulin superfamily containing leucine-rich SEQID NOS: 7229-7232 repeat

ISLR2 Immunoglobulin superfamily containing leucine-rich SEQID NOS: 7233-7242 repeat 2

ISM1 Isthmin 1, angiogenesis inhibitor SEQID NO: 7243

ISM2 Isthmin 2 SEQID NOS: 7244-7249

ITGA4 Integrin, alpha 4 (antigen CD49D, alpha 4 subunit of SEQID NOS: 7250-7252

VLA-4 receptor)

ITGA9 Integrin, alpha 9 SEQID NOS: 7253 -7255

ITGAL Integrin, alpha L (antigen CDllA (pl80), lymphocyte SEQID NOS: 7256-7265 function-associated antigen 1; alpha polypeptide)

ITGAX Integrin, alpha X (complement component 3 SEQID NOS: 7266-7268 receptor 4 subunit)

ITGB1 Integrin, beta 1 (fibronectin receptor, beta SEQID NOS: 7269-7284 polypeptide, antigen CD29 includes MDF2, MSK12)

ITGB2 Integrin, beta 2 (complement component 3 receptor SEQID NOS: 7285-7301

3 and 4 subunit)

ITGB3 Integrin, beta 3 (platelet glycoprotein Ilia, antigen SEQID NOS: 7302-7304

CD61)

ITGB7 Integrin, beta 7 SEQID NOS: 7305-7312

ITGBL1 Integrin, beta-like 1 (with EGF-like repeat domains) SEQID NOS: 7313 -7318 mm Inter-alpha-trypsin inhibitor heavy chain 1 SEQID NOS: 7319-7324

ITIH2 Inter-alpha-trypsin inhibitor heavy chain 2 SEQID NOS: 7325 -7327 ITIH3 Inter-alpha-trypsin inhibitor heavy chain 3 SEQID NOS: 7328-7330

ITIH4 Inter-alpha-trypsin inhibitor heavy chain family, SEQID NOS: 7331 -7334 member 4

ITIH5 Inter-alpha-trypsin inhibitor heavy chain family, SEQID NOS: 7335 -7338 member 5

ITIH6 Inter-alpha-trypsin inhibitor heavy chain family, SEQID NO: 7339

member 6

ITLN1 Intelectin 1 (galactofuranose binding) SEQID NO: 7340

ITLN2 Intelectin 2 SEQID NO: 7341

IZUM01R IZUMOl receptor, JUNO SEQID NOS: 7342-7343

IZUM04 IZUMO family member 4 SEQID NOS: 7344-7350

AMICA1 Adhesion molecule, interacts with CXADR antigen 1 SEQID NOS: 7351 -7359

JCHAIN Joining chain of multimeric IgA and IgM SEQID NOS: 7360-7365

JMJD8 Jumonji domain containing 8 SEQID NOS: 7366-7370

JSRP1 Junctional sarcoplasmic reticulum protein 1 SEQID NO: 7371

KANSL2 KAT8 regulatory NSL complex subunit 2 SEQID NOS: 7372-7382

KAZALD1 Kazal-type serine peptidase inhibitor domain 1 SEQID NO: 7383

KCNIP3 Kv channel interacting protein 3, calsenilin SEQID NOS: 7384-7386

KCNK7 Potassium channel, two pore domain subfamily K, SEQID NOS: 7387-7392 member 7

KCNN4 Potassium channel, calcium activated SEQID NOS: 7393-7398 intermediate/small conductance subfamily N alpha,

member 4

KCNU1 Potassium channel, subfamily U, member 1 SEQID NOS: 7399-7403

KCP Kielin/chordin-like protein SEQID NOS: 7404-7407

KDELC1 KDEL (Lys-Asp-Glu-Leu) containing 1 SEQID NO: 7408

KDELC2 KDEL (Lys-Asp-Glu-Leu) containing 2 SEQID NOS: 7409-7412

KDM1A Lysine (K)-specific demethylase 1A SEQID NOS: 7413-7416

KDM3B Lysine (K)-specific demethylase 3B SEQID NOS: 7417-7420

KDM6A Lysine (K)-specific demethylase 6A SEQID NOS: 7421-7430

KDM7A Lysine (K)-specific demethylase 7A SEQID NOS: 7431-7432

KDSR 3-ketodihydrosphingosine reductase SEQID NOS: 7433-7439

KERA Keratocan SEQID NO: 7440

KIAA0100 KIAA0100 SEQID NOS: 7441-7446

KIAA0319 KIAA0319 SEQID NOS: 7447-7452

KIAA1324 KIAA1324 SEQID NOS: 7453-7461

KIFC2 Kinesin family member C2 SEQID NOS: 7462-7464

KIR2DL4 Killer cell immunoglobulin-like receptor, two SEQID NOS: 7465-7471 domains, long cytoplasmic tail, 4

KIR3DX1 Killer cell immunoglobulin-like receptor, three SEQID NOS: 7472-7476 domains, XI

KIRREL2 Kin of IRRE like 2 (Drosophila) SEQID NOS: 7477-7481

KISS1 KiSS-1 metastasis-suppressor SEQID NOS: 7482-7483

KLHL11 Kelch-like family member 11 SEQID NO: 7484

KLHL22 Kelch-like family member 22 SEQID NOS: 7485-7491

KLK1 Kallikrein 1 SEQID NOS: 7492-7493

KLK10 Kallikrein-related peptidase 10 SEQID NOS: 7494-7498

KLK11 Kallikrein-related peptidase 11 SEQID NOS: 7499-7507 KLK12 Kallikrein-related peptidase 12 SEQID NOS: 7508-7514

KLK13 Kallikrein-related peptidase 13 SEQID NOS: 7515 -7523

KLK14 Kallikrein-related peptidase 14 SEQID NOS: 7524-7525

KLK15 Kallikrein-related peptidase 15 SEQID NOS: 7526-7530

KLK2 Kallikrein-related peptidase 2 SEQID NOS: 7531-7543

KLK3 Kallikrein-related peptidase 3 SEQID NOS: 7544-7555

KLK4 Kallikrein-related peptidase 4 SEQID NOS: 7556-7560

KLK5 Kallikrein-related peptidase 5 SEQID NOS: 7561-7564

KLK6 Kallikrein-related peptidase 6 SEQID NOS: 7565-7571

KLK7 Kallikrein-related peptidase 7 SEQID NOS: 7572-7576

KLK8 Kallikrein-related peptidase 8 SEQID NOS: 7577-7584

KLK9 Kallikrein-related peptidase 9 SEQID NOS: 7585-7586

KLKB1 Kallikrein B, plasma (Fletcher factor) 1 SEQID NOS: 7587-7591

SETD8 SET domain containing (lysine methyltransferase) 8 SEQID NOS: 7592-7595

KNDC1 Kinase non-catalytic C-lobe domain (KIND) SEQID NOS: 7596-7597 containing 1

KNG1 Kininogen 1 SEQID NOS: 7598-7602

KRBA2 KRAB-A domain containing 2 SEQID NOS: 7603-7606

KREMEN2 Kringle containing transmembrane protein 2 SEQID NOS: 7607-7612

KRTDAP Keratinocyte differentiation-associated protein SEQID NOS: 7613-7614

L1CAM LI cell adhesion molecule SEQID NOS: 7615-7624

L3MBTL2 L(3)mbt-like 2 (Drosophila) SEQID NOS: 7625-7629

LACRT Lacritin SEQID NOS: 7630-7632

LACTB Lactamase, beta SEQID NOS: 7633-7635

LAG 3 Lymphocyte-activation gene 3 SEQID NOS: 7636-7637

LAIR2 Leukocyte-associated immunoglobulin-like receptor SEQID NOS: 7638-7641

2

LALBA Lactalbumin, alpha- SEQID NOS: 7642-7643

LAMA1 Laminin, alpha 1 SEQID NOS: 7644-7645

LAMA2 Laminin, alpha 2 SEQID NOS: 7646-7649

LAMA3 Laminin, alpha 3 SEQID NOS: 7650-7659

LAMA4 Laminin, alpha 4 SEQID NOS: 7660-7674

LAMA5 Laminin, alpha 5 SEQID NOS: 7675-7677

LAMB1 Laminin, beta 1 SEQID NOS: 7678-7682

LAMB2 Laminin, beta 2 (laminin S) SEQID NOS: 7683-7685

LAMB3 Laminin, beta 3 SEQID NOS: 7686-7690

LAMB4 Laminin, beta 4 SEQID NOS: 7691-7694

LAMC1 Laminin, gamma 1 (formerly LAMB2) SEQID NOS: 7695-7696

LAMC2 Laminin, gamma 2 SEQID NOS: 7697-7698

LAMC3 Laminin, gamma 3 SEQID NOS: 7699-7700

LAMP3 Lysosomal-associated membrane protein 3 SEQID NOS: 7701-7704

GYLTL1B Glycosyltransferase-like IB SEQID NOS: 7705-7710

LAT Linker for activation of T cells SEQID NOS: 7711-7720

LAT2 Linker for activation of T cells family, member 2 SEQID NOS: 7721-7729

LBP Lipopolysaccharide binding protein SEQID NO: 7730

LCAT Lecithin-cholesterol acyltransferase SEQID NOS: 7731-7737

LCN1 Lipocalin 1 SEQID NOS: 7738-7739 LCN10 Lipocalin 10 SEQID NOS: 7740-7745

LCN12 Lipocalin 12 SEQID NOS: 7746-7748

LCN15 Lipocalin 15 SEQID NO: 7749

LCN2 Lipocalin 2 SEQID NOS: 7750-7752

LCN6 Lipocalin 6 SEQID NOS: 7753-7754

LCN8 Lipocalin 8 SEQID NOS: 7755-7756

LCN9 Lipocalin 9 SEQID NOS: 7757-7758

LCORL Ligand dependent nuclear receptor corepressor-like SEQID NOS: 7759-7764

LDLR Low density lipoprotein receptor SEQID NOS: 7765-7773

LDLRAD2 Low density lipoprotein receptor class A domain SEQID NOS: 7774-7775 containing 2

LEAP2 Liver expressed antimicrobial peptide 2 SEQID NO: 7776

LECT2 Leukocyte cell-derived chemotaxin 2 SEQID NOS: 7777-7780

LEFTY1 Left-right determination factor 1 SEQID NOS: 7781-7782

LEFTY2 Left-right determination factor 2 SEQID NOS: 7783-7784

LEP Leptin SEQID NO: 7785

LFNG LFNG O-fucosylpeptide 3-beta-N- SEQID NOS: 7786-7791 acetylglucosaminyltransferase

LGALS3BP Lectin, galactoside-binding, soluble, 3 binding SEQID NOS: 7792-7806 protein

LGI1 Leucine-rich, glioma inactivated 1 SEQID NOS: 7807-7825

LGI2 Leucine-rich repeat LGI family, member 2 SEQID NOS: 7826-7827

LGI3 Leucine-rich repeat LGI family, member 3 SEQID NOS: 7828-7831

LGI4 Leucine-rich repeat LGI family, member 4 SEQID NOS: 7832-7835

LGMN Legumain SEQID NOS: 7836-7849

LGR4 Leucine-rich repeat containing G protein-coupled SEQID NOS: 7850-7852 receptor 4

LHB Luteinizing hormone beta polypeptide SEQID NO: 7853

LHCGR Luteinizing hormone/choriogonadotropin receptor SEQID NOS: 7854-7858

LIF Leukemia inhibitory factor SEQID NOS: 7859-7860

LIFR Leukemia inhibitory factor receptor alpha SEQID NOS: 7861-7865

LILRAl Leukocyte immunoglobulin-like receptor, subfamily SEQID NOS: 7866-7867

A (with TM domain), member 1

LILRA2 Leukocyte immunoglobulin-like receptor, subfamily SEQID NOS: 7868-7874

A (with TM domain), member 2

LILRB3 Leukocyte immunoglobulin-like receptor, subfamily SEQID NOS: 7875-7879

B (with TM and ITIM domains), member 3

LIMEl Lck interacting transmembrane adaptor 1 SEQID NOS: 7880-7885

LINGOl Leucine rich repeat and Ig domain containing 1 SEQID NOS: 7886-7896

LI PA Lipase A, lysosomal acid, cholesterol esterase SEQID NOS: 7897-7901

UPC Lipase, hepatic SEQID NOS: 7902-7905

LIPF Lipase, gastric SEQID NOS: 7906-7909

LIPG Lipase, endothelial SEQID NOS: 7910-7915

LIPH Lipase, member H SEQID NOS: 7916-7920

LIPK Lipase, family member K SEQID NO: 7921

LIPM Lipase, family member M SEQID NOS: 7922-7923

LIPN Lipase, family member N SEQID NO: 7924 LMAN2 Lectin, mannose-binding 2 SEQID NOS: 7925-7929

LMNTD1 Lamin tail domain containing 1 SEQID NOS: 7930-7940

LNX1 Ligand of numb-protein X 1, E3 ubiquitin protein SEQID NOS: 7941-7947 ligase

LOX Lysyl oxidase SEQID NOS: 7948-7950

LOXL1 Lysyl oxidase-like 1 SEQID NOS: 7951-7952

LOXL2 Lysyl oxidase-like 2 SEQID NOS: 7953-7961

LOXL3 Lysyl oxidase-like 3 SEQID NOS: 7962-7968

LOXL4 Lysyl oxidase-like 4 SEQID NO: 7969

LPA Lipoprotein, Lp(a) SEQID NOS: 7970-7972

LPL Lipoprotein lipase SEQID NOS: 7973-7977

LPO Lactoperoxidase SEQID NOS: 7978-7984

LRAT Lecithin retinol acyltransferase SEQID NOS: 7985-7987

(phosphatidylcholine— retinol O-acyltransferase)

LRCH3 Leucine-rich repeats and calponin homology (CH) SEQID NOS: 7988-7996 domain containing 3

LRCOL1 Leucine rich colipase-like 1 SEQID NOS: 7997-8000

LRFN4 Leucine rich repeat and fibronectin type III domain SEQID NOS: 8001-8002 containing 4

LRFN5 Leucine rich repeat and fibronectin type III domain SEQID NOS: 8003-8005 containing 5

LRG1 Leucine-rich alpha-2-glycoprotein 1 SEQID NO: 8006

LRP1 Low density lipoprotein receptor-related protein 1 SEQID NOS: 8007-8012

LRP11 Low density lipoprotein receptor-related protein 11 SEQID NOS: 8013-8014

LRP1B Low density lipoprotein receptor-related protein IB SEQID NOS: 8015-8018

LRP2 Low density lipoprotein receptor-related protein 2 SEQID NOS: 8019-8020

LRP4 Low density lipoprotein receptor-related protein 4 SEQID NOS: 8021-8022

LRPAP1 Low density lipoprotein receptor-related protein SEQID NOS: 8023-8024 associated protein 1

LRRC17 Leucine rich repeat containing 17 SEQID NOS: 8025-8027

LRRC32 Leucine rich repeat containing 32 SEQID NOS: 8028-8031

LRRC3B Leucine rich repeat containing 3B SEQID NOS: 8032-8036

LRRC4B Leucine rich repeat containing 4B SEQID NOS: 8037-8039

LRRC70 Leucine rich repeat containing 70 SEQID NOS: 8040-8041

LRRN3 Leucine rich repeat neuronal 3 SEQID NOS: 8042-8045

LRRTM1 Leucine rich repeat transmembrane neuronal 1 SEQID NOS: 8046-8052

LRRTM2 Leucine rich repeat transmembrane neuronal 2 SEQID NOS: 8053-8055

LRRTM4 Leucine rich repeat transmembrane neuronal 4 SEQID NOS: 8056-8061

LRTM2 Leucine-rich repeats and transmembrane domains 2 SEQID NOS: 8062-8066

LSR Lipolysis stimulated lipoprotein receptor SEQID NOS: 8067-8077

LST1 Leukocyte specific transcript 1 SEQID NOS: 8078-8095

LTA Lymphotoxin alpha SEQID NOS: 8096-8097

LTBP1 Latent transforming growth factor beta binding SEQID NOS: 8098-8107 protein 1

LTBP2 Latent transforming growth factor beta binding SEQID NOS: 8108-8111 protein 2

LTBP3 Latent transforming growth factor beta binding SEQID NOS: 8112-8124 protein 3 LTBP4 Latent transforming growth factor beta binding SEQID NOS: 8125-8140 protein 4

LTBR Lymphotoxin beta receptor (TNFR superfamily, SEQID NOS: 8141-8146 member 3)

LTF Lactotransferrin SEQID NOS: 8147-8151

LTK Leukocyte receptor tyrosine kinase SEQID NOS: 8152-8155

LUM Lumican SEQID NO: 8156

LUZP2 Leucine zipper protein 2 SEQID NOS: 8157-8160

LVRN Laeverin SEQID NOS: 8161-8166

LY6E Lymphocyte antigen 6 complex, locus E SEQID NOS: 8167-8180

LY6G5B Lymphocyte antigen 6 complex, locus G5B SEQID NOS: 8181-8182

LY6G6D Lymphocyte antigen 6 complex, locus G6D SEQID NOS: 8183-8184

LY6G6E Lymphocyte antigen 6 complex, locus G6E SEQID NOS: 8185-8188

(pseudogene)

LY6H Lymphocyte antigen 6 complex, locus H SEQID NOS: 8189-8192

LY6K Lymphocyte antigen 6 complex, locus K SEQID NOS: 8193-8196

RP11- SEQID NO: 8197 520P18.5

LY86 Lymphocyte antigen 86 SEQID NOS: 8198-8199

LY96 Lymphocyte antigen 96 SEQID NOS: 8200-8201

LYG1 Lysozyme G-like 1 SEQID NOS: 8202-8203

LYG2 Lysozyme G-like 2 SEQID NOS: 8204-8209

LYNX1 Ly6/neurotoxin 1 SEQID NOS: 8210-8214

LYPD1 LY6/PLAUR domain containing 1 SEQID NOS: 8215 -8217

LYPD2 LY6/PLAUR domain containing 2 SEQID NO: 8218

LYPD4 LY6/PLAUR domain containing 4 SEQID NOS: 8219-8221

LYPD6 LY6/PLAUR domain containing 6 SEQID NOS: 8222 -8226

LYPD6B LY6/PLAUR domain containing 6B SEQID NOS: 8227-8233

LYPD8 LY6/PLAUR domain containing 8 SEQID NOS: 8234-8235

LYZ Lysozyme SEQID NOS: 8236-8238

LYZL4 Lysozyme-like 4 SEQID NOS: 8239-8240

LYZL6 Lysozyme-like 6 SEQID NOS: 8241-8243

M6PR Mannose-6-phosphate receptor (cation dependent) SEQID NOS: 8244-8254

MAD1L1 MAD1 mitotic arrest deficient-like 1 (yeast) SEQID NOS: 8255 -8267

MAG Myelin associated glycoprotein SEQID NOS: 8268-8273

MAGT1 Magnesium transporter 1 SEQID NOS: 8274-8277

MALSU1 Mitochondrial assembly of ribosomal large subunit 1 SEQID NO: 8278

MAMDC2 MAM domain containing 2 SEQID NO: 8279

MAN2B1 Mannosidase, alpha, class 2B, member 1 SEQID NOS: 8280-8285

MAN2B2 Mannosidase, alpha, class 2B, member 2 SEQID NOS: 8286-8288

MANBA Mannosidase, beta A, lysosomal SEQID NOS: 8289-8302

MANEAL Mannosidase, endo-alpha-like SEQID NOS: 8303-8307

MANF Mesencephalic astrocyte-derived neurotrophic SEQID NOS: 8308-8309 factor

MANSC1 MANSC domain containing 1 SEQID NOS: 8310-8313

MAP3K9 Mitogen-activated protein kinase 9 SEQID NOS: 8314-8319 MAS PI Mannan-binding lectin serine peptidase 1 (C4/C2 SEQID NOS: 8320-8327 activating component of Ra-reactive factor)

MASP2 Mannan-binding lectin serine peptidase 2 SEQID NOS: 8328-8329

MATN1 Matrilin 1, cartilage matrix protein SEQID NO: 8330

MATN2 Matrilin 2 SEQID NOS: 8331-8343

MATN3 Matrilin 3 SEQID NOS: 8344-8345

MATN4 Matrilin 4 SEQID NOS: 8346-8350

MATR3 Matrin 3 SEQID NOS: 8351-8378

MAU2 MAU2 sister chromatid cohesion factor SEQID NOS: 8379-8381

MAZ MYC-associated zinc finger protein (purine-binding SEQID NOS: 8382-8396 transcription factor)

MBD6 Methyl-CpG binding domain protein 6 SEQID NOS: 8397-8408

MBL2 Mannose-binding lectin (protein C) 2, soluble SEQID NO: 8409

MBNL1 Muscleblind-like splicing regulator 1 SEQID NOS: 8410-8428

MCCC1 Methylcrotonoyl-CoA carboxylase 1 (alpha) SEQID NOS: 8429-8440

MCCD1 Mitochondrial coiled-coil domain 1 SEQID NO: 8441

MCEE Methylmalonyl CoA epimerase SEQID NOS: 8442-8445

MCF2L MCF.2 cell line derived transforming sequence-like SEQID NOS: 8446-8467

MCFD2 Multiple coagulation factor deficiency 2 SEQID NOS: 8468-8479

MDFIC MyoD family inhibitor domain containing SEQID NOS: 8480-8487

MDGA1 MAM domain containing SEQID NOS: 8488-8493 glycosylphosphatidylinositol anchor 1

MDK Midkine (neurite growth-promoting factor 2) SEQID NOS: 8494-8503

MED20 Mediator complex subunit 20 SEQID NOS: 8504-8508

MEGF10 Multiple EGF-like-domains 10 SEQID NOS: 8509-8512

MEGF6 Multiple EGF-like-domains 6 SEQID NOS: 8513 -8516

MEI1 Meiotic double-stranded break formation protein 1 SEQID NOS: 8517-8520

MEI4 Meiotic double-stranded break formation protein 4 SEQID NO: 8521

MEIS1 Meis homeobox 1 SEQID NOS: 8522 -8527

MEIS3 Meis homeobox 3 SEQID NOS: 8528-8537

MFI2 Antigen p97 (melanoma associated) identified by SEQID NOS: 8538-8540 monoclonal antibodies 133.2 and 96.5

MEPE Matrix extracellular phosphoglycoprotein SEQID NOS: 8541-8547

MESDC2 Mesoderm development candidate 2 SEQID NOS: 8548-8552

MEST Mesoderm specific transcript SEQID NOS: 8553 -8566

MET MET proto-oncogene, receptor tyrosine kinase SEQID NOS: 8567-8572

METRN Meteorin, glial cell differentiation regulator SEQID NOS: 8573-8577

METRNL Meteorin, glial cell differentiation regulator-like SEQID NOS: 8578-8581

METTL17 Methyltransferase like 17 SEQID NOS: 8582-8592

METTL24 Methyltransferase like 24 SEQID NO: 8593

METTL7B Methyltransferase like 7B SEQID NOS: 8594-8595

METTL9 Methyltransferase like 9 SEQID NOS: 8596-8604

MEX3C Mex-3 RNA binding family member C SEQID NOS: 8605-8607

MFAP2 Microfibrillar-associated protein 2 SEQID NOS: 8608-8609

MFAP3 Microfibrillar-associated protein 3 SEQID NOS: 8610-8614

MFAP3L Microfibrillar-associated protein 3-like SEQID NOS: 8615-8624

MFAP4 Microfibrillar-associated protein 4 SEQID NOS: 8625-8627 MFAP5 Microfibrillar associated protein 5 SEQID NOS: 8628-8638

MFGE8 Milk fat globule-EGF factor 8 protein SEQID NOS: 8639-8645

MFNG MFNG O-fucosylpeptide 3-beta-N- SEQID NOS: 8646-8653 acetylglucosaminyltransferase

MGA MGA, MAX dimerization protein SEQID NOS: 8654-8662

MGAT2 Mannosyl (alpha-l,6-)-glycoprotein beta-l,2-N- SEQID NO: 8663

acetylglucosaminyltransferase

MGAT3 Mannosyl (beta-l,4-)-glycoprotein beta-l,4-N- SEQID NOS: 8664-8666 acetylglucosaminyltransferase

MGAT4A Mannosyl (alpha-l,3-)-glycoprotein beta-l,4-N- SEQID NOS: 8667-8671 acetylglucosaminyltransferase, isozyme A

MGAT4B Mannosyl (alpha-l,3-)-glycoprotein beta-l,4-N- SEQID NOS: 8672-8682 acetylglucosaminyltransferase, isozyme B

MGAT4D MGAT4 family, member D SEQID NOS: 8683-8688

MGLL Monoglyceride lipase SEQID NOS: 8689-8698

MGP Matrix Gla protein SEQID NOS: 8699-8701

MGST2 Microsomal glutathione S-transferase 2 SEQID NOS: 8702-8705

MIA Melanoma inhibitory activity SEQID NOS: 8706-8711

MIA2 Melanoma inhibitory activity 2 SEQID NO: 8712

MIA3 Melanoma inhibitory activity family, member 3 SEQID NOS: 8713-8717

MICUl Mitochondrial calcium uptake 1 SEQID NOS: 8718-8727

MIER1 Mesoderm induction early response 1, SEQID NOS: 8728-8736 transcriptional regulator

MIN0S1- MINOS1-NBL1 readthrough SEQID NOS: 8737-8739 NBL1

MINPP1 Multiple inositol-polyphosphate phosphatase 1 SEQID NOS: 8740-8742

MLEC Malectin SEQID NOS: 8743-8746

MLN Motilin SEQID NOS: 8747-8749

MLXIP MLX interacting protein SEQID NOS: 8750-8755

MLXIPL MLX interacting protein-like SEQID NOS: 8756-8763

MMP1 Matrix metallopeptidase 1 SEQID NO: 8764

MMP10 Matrix metallopeptidase 10 SEQID NOS: 8765-8766

MMP11 Matrix metallopeptidase 11 SEQID NOS: 8767-8770

MMP12 Matrix metallopeptidase 12 SEQID NO: 8771

MMP13 Matrix metallopeptidase 13 SEQID NOS: 8772-8774

MMP14 Matrix metallopeptidase 14 (membrane-inserted) SEQID NOS: 8775-8777

MMP17 Matrix metallopeptidase 17 (membrane-inserted) SEQID NOS: 8778-8785

MMP19 Matrix metallopeptidase 19 SEQID NOS: 8786-8791

MMP2 Matrix metallopeptidase 2 SEQID NOS: 8792-8799

MMP20 Matrix metallopeptidase 20 SEQID NO: 8800

MMP21 Matrix metallopeptidase 21 SEQID NO: 8801

MMP25 Matrix metallopeptidase 25 SEQID NOS: 8802-8803

MMP26 Matrix metallopeptidase 26 SEQID NOS: 8804-8805

MMP27 Matrix metallopeptidase 27 SEQID NO: 8806

MMP28 Matrix metallopeptidase 28 SEQID NOS: 8807-8812

MMP3 Matrix metallopeptidase 3 SEQID NOS: 8813-8815

MMP7 Matrix metallopeptidase 7 SEQID NO: 8816 MMP8 Matrix metallopeptidase 8 SEQID NOS: 8817-8822

MMP9 Matrix metallopeptidase 9 SEQID NO: 8823

MMRN1 Multimerin 1 SEQID NOS: 8824-8826

MMRN2 Multimerin 2 SEQID NOS: 8827-8831

MOXD1 Monooxygenase, DBH-like 1 SEQID NOS: 8832-8834

C6orf25 Chromosome 6 open reading frame 25 SEQID NOS: 8835-8842

MPO Myeloperoxidase SEQID NOS: 8843-8844

MPPED1 Metallophosphoesterase domain containing 1 SEQID NOS: 8845-8848

MPZL1 Myelin protein zero-like 1 SEQID NOS: 8849-8853

MR1 Major histocompatibility complex, class l-related SEQID NOS: 8854-8859

MRPL2 Mitochondrial ribosomal protein L2 SEQID NOS: 8860-8864

MRPL21 Mitochondrial ribosomal protein L21 SEQID NOS: 8865-8871

MRPL22 Mitochondrial ribosomal protein L22 SEQID NOS: 8872-8876

MRPL24 Mitochondrial ribosomal protein L24 SEQID NOS: 8877-8881

MRPL27 Mitochondrial ribosomal protein L27 SEQID NOS: 8882-8887

MRPL32 Mitochondrial ribosomal protein L32 SEQID NOS: 8888-8890

MRPL34 Mitochondrial ribosomal protein L34 SEQID NOS: 8891-8895

MRPL35 Mitochondrial ribosomal protein L35 SEQID NOS: 8896-8899

MRPL52 Mitochondrial ribosomal protein L52 SEQID NOS: 8900-8910

MRPL55 Mitochondrial ribosomal protein L55 SEQID NOS: 8911-8936

MRPS14 Mitochondrial ribosomal protein S14 SEQID NOS: 8937-8938

MRPS22 Mitochondrial ribosomal protein S22 SEQID NOS: 8939-8947

MRPS28 Mitochondrial ribosomal protein S28 SEQID NOS: 8948-8955

MS4A14 Membrane-spanning 4-domains, subfamily A, SEQID NOS: 8956-8966 member 14

MS4A3 Membrane-spanning 4-domains, subfamily A, SEQID NOS: 8967-8971 member 3 (hematopoietic cell-specific)

MSH3 MutS homolog 3 SEQID NO: 8972

MSH5 MutS homolog 5 SEQID NOS: 8973-8984

MSLN Mesothelin SEQID NOS: 8985-8992

MSMB Microseminoprotein, beta- SEQID NOS: 8993-8994

MSRA Methionine sulfoxide reductase A SEQID NOS: 8995-9002

MSRB2 Methionine sulfoxide reductase B2 SEQID NOS: 9003-9004

MSRB3 Methionine sulfoxide reductase B3 SEQID NOS: 9005-9018

MST1 Macrophage stimulating 1 SEQID NOS: 9019-9020

MSTN Myostatin SEQID NO: 9021

MT1G Metallothionein 1G SEQID NOS: 9022-9025

MTHFD2 Methylenetetrahydrofolate dehydrogenase (NADP+ SEQID NOS: 9026-9030 dependent) 2, methenyltetrahydrofolate

cyclohydrolase

MTMR14 Myotubularin related protein 14 SEQID NOS: 9031-9041

MTRNR2L11 MT-RNR2-like 11 (pseudogene) SEQID NO: 9042

MTRR 5-methyltetrahydrofolate-homocysteine SEQID NOS: 9043-9055 methyltransferase reductase

MTTP Microsomal triglyceride transfer protein SEQID NOS: 9056-9066

MTX2 Metaxin 2 SEQID NOS: 9067-9071

MUC1 Mucin 1, cell surface associated SEQID NOS: 9072-9097 MUC13 Mucin 13, cell surface associated SEQID NOS: 9098-9099

MUC20 Mucin 20, cell surface associated SEQID NOS: 9100-9104

MUC3A Mucin 3A, cell surface associated SEQID NOS: 9105-9107

MUC5AC Mucin 5AC, oligomeric mucus/gel-forming SEQID NO: 9108

MUC5B Mucin 5B, oligomeric mucus/gel-forming SEQID NOS: 9109-9110

MUC6 Mucin 6, oligomeric mucus/gel-forming SEQID NOS: 9111-9114

MUC7 Mucin 7, secreted SEQID NOS: 9115-9118

MUCL1 Mucin-like 1 SEQID NOS: 9119-9121

MXRA5 Matrix-remodelling associated 5 SEQID NO: 9122

MXRA7 Matrix-remodelling associated 7 SEQID NOS: 9123-9129

MYDGF Myeloid-derived growth factor SEQID NOS: 9130-9132

MYL1 Myosin, light chain 1, alkali; skeletal, fast SEQID NOS: 9133-9134

MYOC Myocilin, trabecular meshwork inducible SEQID NOS: 9135-9136 glucocorticoid response

MYRFL Myelin regulatory factor-like SEQID NOS: 9137-9141

MZB1 Marginal zone B and Bl cell-specific protein SEQID NOS: 9142-9146

N4BP2L2 NEDD4 binding protein 2-like 2 SEQID NOS: 9147-9152

NAA38 N(alpha)-acetyltransferase 38, NatC auxiliary subunit SEQID NOS: 9153-9158

NAAA N-acylethanolamine acid amidase SEQID NOS: 9159-9164

NAGA N-acetylgalactosaminidase, alpha- SEQID NOS: 9165-9167

NAGLU N-acetylglucosaminidase, alpha SEQID NOS: 9168-9172

NAGS N-acetylglutamate synthase SEQID NOS: 9173-9174

NAPSA Napsin A aspartic peptidase SEQID NOS: 9175-9177

CARKD Carbohydrate kinase domain containing SEQID NOS: 9178-9179

APOA1BP Apolipoprotein A-l binding protein SEQID NOS: 9180-9182

NBL1 Neuroblastoma 1, DAN family BMP antagonist SEQID NOS: 9183-9196

NCAM1 Neural cell adhesion molecule 1 SEQID NOS: 9197-9216

NCAN Neurocan SEQID NOS: 9217-9218

NCBP2-AS2 NCBP2 antisense RNA 2 (head to head) SEQID NO: 9219

NCSTN Nicastrin SEQID NOS: 9220-9229

NDNF Neuron-derived neurotrophic factor SEQID NOS: 9230-9232

NDP Norrie disease (pseudoglioma) SEQID NOS: 9233 -9235

NDUFA10 NADH dehydrogenase (ubiquinone) 1 alpha SEQID NOS: 9236-9245 subcomplex, 10, 42kDa

NDUFB5 NADH dehydrogenase (ubiquinone) 1 beta SEQID NOS: 9246-9254 subcomplex, 5, 16kDa

NDUFS8 NADH dehydrogenase (ubiquinone) Fe-S protein 8, SEQID NOS: 9255 -9264

23kDa (NADH-coenzyme Q reductase)

NDUFV1 NADH dehydrogenase (ubiquinone) flavoprotein 1, SEQID NOS: 9265-9278

51kDa

NECAB3 N-terminal EF-hand calcium binding protein 3 SEQID NOS: 9279-9288

PVRL1 Poliovirus receptor-related 1 (herpesvirus entry SEQID NOS: 9289-9291 mediator C)

NELL1 Neural EGFL like 1 SEQID NOS: 9292-9295

NELL2 Neural EGFL like 2 SEQID NOS: 9296-9310

NENF Neudesin neurotrophic factor SEQID NO: 9311

NETOl Neuropilin (NRP) and tolloid (TLL)-like 1 SEQID NOS: 9312 -9316 NFASC Neurofascin SEQID NOS: 9317-9331

NFE2L1 Nuclear factor, erythroid 2-like 1 SEQID NOS: 9332 -9350

NFE2L3 Nuclear factor, erythroid 2-like 3 SEQID NOS: 9351 -9352

NGEF Neuronal guanine nucleotide exchange factor SEQID NOS: 9353 -9358

NGF Nerve growth factor (beta polypeptide) SEQID NO: 9359

NGLY1 N-glycanase 1 SEQID NOS: 9360-9366

NGRN Neugrin, neurite outgrowth associated SEQID NOS: 9367-9368

NHLRC3 NHL repeat containing 3 SEQID NOS: 9369-9371

NIDI Nidogen 1 SEQID NOS: 9372 -9373

NID2 Nidogen 2 (osteonidogen) SEQID NOS: 9374-9376

NKG7 Natural killer cell granule protein 7 SEQID NOS: 9377-9381

NLGN3 Neuroligin 3 SEQID NOS: 9382 -9386

NLGN4Y Neuroligin 4, Y-linked SEQID NOS: 9387-9393

NLRP5 NLR family, pyrin domain containing 5 SEQID NOS: 9394-9396

NMB Neuromedin B SEQID NOS: 9397-9398

NME1 NME/NM23 nucleoside diphosphate kinase 1 SEQID NOS: 9399-9405

NME1- NME1-NME2 readthrough SEQID NOS: 9406-9408 NME2

NME3 NME/NM23 nucleoside diphosphate kinase 3 SEQID NOS: 9409-9413

NMS Neuromedin S SEQID NO: 9414

NMU Neuromedin U SEQID NOS: 9415 -9418

N0A1 Nitric oxide associated 1 SEQID NO: 9419

NODAL Nodal growth differentiation factor SEQID NOS: 9420-9421

NOG Noggin SEQID NO: 9422

NOM03 NODAL modulator 3 SEQID NOS: 9423 -9429

NOS1AP Nitric oxide synthase 1 (neuronal) adaptor protein SEQID NOS: 9430-9434

NOTCH3 Notch 3 SEQID NOS: 9435 -9438

NOTUM Notum pectinacetylesterase homolog (Drosophila) SEQID NOS: 9439-9441

NOV Nephroblastoma overexpressed SEQID NO: 9442

NPB Neuropeptide B SEQID NOS: 9443 -9444

NPC2 Niemann-Pick disease, type C2 SEQID NOS: 9445 -9453

NPFF Neuropeptide FF-amide peptide precursor SEQID NO: 9454

NPFFR2 Neuropeptide FF receptor 2 SEQID NOS: 9455 -9458

NPHS1 Nephrosis 1, congenital, Finnish type (nephrin) SEQID NOS: 9459-9460

NPNT Nephronectin SEQID NOS: 9461 -9471

NPPA Natriuretic peptide A SEQID NOS: 9472 -9474

NPPB Natriuretic peptide B SEQID NO: 9475

NPPC Natriuretic peptide C SEQID NOS: 9476-9477

NPS Neuropeptide S SEQID NO: 9478

NPTX1 Neuronal pentraxin 1 SEQID NO: 9479

NPTX2 Neuronal pentraxin II SEQID NO: 9480

NPTXR Neuronal pentraxin receptor SEQID NOS: 9481 -9482

NPVF Neuropeptide VF precursor SEQID NO: 9483

NPW Neuropeptide W SEQID NOS: 9484-9486

NPY Neuropeptide Y SEQID NOS: 9487-9489

NQ02 NAD(P)H dehydrogenase, quinone 2 SEQID NOS: 9490-9498 NRCAM Neuronal cell adhesion molecule SEQID NOS: 9499-9511

NRG1 Neuregulin 1 SEQID NOS: 9512 -9529

NRN1L Neuritin 1-like SEQID NOS: 9530-9532

NRP1 Neuropilin 1 SEQID NOS: 9533-9546

NRP2 Neuropilin 2 SEQID NOS: 9547-9553

NRTN Neurturin SEQID NO: 9554

NRXN1 Neurexin 1 SEQID NOS: 9555 -9585

NRXN2 Neurexin 2 SEQID NOS: 9586-9594

NT5C3A 5'-nucleotidase, cytosolic IIIA SEQID NOS: 9595-9605

NT5DC3 5'-nucleotidase domain containing 3 SEQID NOS: 9606-9608

NT5E 5'-nucleotidase, ecto (CD73) SEQID NOS: 9609-9613

NTF3 Neurotrophin 3 SEQID NOS: 9614-9615

NTF4 Neurotrophin 4 SEQID NOS: 9616-9617

NTM Neurotrimin SEQID NOS: 9618-9627

NTN1 Netrin 1 SEQID NOS: 9628-9629

NTN3 Netrin 3 SEQID NO: 9630

NTN4 Netrin 4 SEQID NOS: 9631-9635

NTN5 Netrin 5 SEQID NOS: 9636-9637

NTNG1 Netrin Gl SEQID NOS: 9638-9644

NTNG2 Netrin G2 SEQID NOS: 9645-9646

NTS Neurotensin SEQID NOS: 9647-9648

NUBPL Nucleotide binding protein-like SEQID NOS: 9649-9655

NUCB1 Nucleobindin 1 SEQID NOS: 9656-9662

NUCB2 Nucleobindin 2 SEQID NOS: 9663-9678

NUDT19 Nudix (nucleoside diphosphate linked moiety X)-type SEQID NO: 9679

motif 19

NUDT9 Nudix (nucleoside diphosphate linked moiety X)-type SEQID NOS: 9680-9684 motif 9

NUP155 Nucleoporin 155kDa SEQID NOS: 9685-9688

NUP214 Nucleoporin 214kDa SEQID NOS: 9689-9700

NUP85 Nucleoporin 85kDa SEQID NOS: 9701-9715

NXPE3 Neurexophilin and PC-esterase domain family, SEQID NOS: 9716-9721 member 3

NXPE4 Neurexophilin and PC-esterase domain family, SEQID NOS: 9722-9723 member 4

NXPH1 Neurexophilin 1 SEQID NOS: 9724-9727

NXPH2 Neurexophilin 2 SEQID NO: 9728

NXPH3 Neurexophilin 3 SEQID NOS: 9729-9730

NXPH4 Neurexophilin 4 SEQID NOS: 9731-9732

NYX Nyctalopin SEQID NOS: 9733-9734

OAF Out at first homolog SEQID NOS: 9735-9736

OBP2A Odorant binding protein 2A SEQID NOS: 9737-9743

OBP2B Odorant binding protein 2B SEQID NOS: 9744-9747

OC90 Otoconin 90 SEQID NO: 9748

OCLN Occludin SEQID NOS: 9749-9751

ODAM Odontogenic, ameloblast asssociated SEQID NOS: 9752-9755

C4orf26 Chromosome 4 open reading frame 26 SEQID NOS: 9756-9759 OGG1 8-oxoguanine DNA glycosylase SEQID NOS: 9760-9773

OGN Osteoglycin SEQID NOS: 9774-9776

OIT3 Oncoprotein induced transcript 3 SEQID NOS: 9777-9778

OLFM1 Olfactomedin 1 SEQID NOS: 9779-9789

OLFM2 Olfactomedin 2 SEQID NOS: 9790-9793

OLFM3 Olfactomedin 3 SEQID NOS: 9794-9796

OLFM4 Olfactomedin 4 SEQID NO: 9797

OLFML1 Olfactomedin-like 1 SEQID NOS: 9798-9801

OLFML2A Olfactomedin-like 2A SEQID NOS: 9802 -9804

OLFML2B Olfactomedin-like 2B SEQID NOS: 9805 -9809

OLFML3 Olfactomedin-like 3 SEQID NOS: 9810-9812

OMD Osteomodulin SEQID NO: 9813

OMG Oligodendrocyte myelin glycoprotein SEQID NO: 9814

OOSP2 Oocyte secreted protein 2 SEQID NOS: 9815 -9816

OPCML Opioid binding protein/cell adhesion molecule-like SEQID NOS: 9817-9821

PROL1 Proline rich, lacrimal 1 SEQID NO: 9822

OPTC Opticin SEQID NOS: 9823 -9824

ORAI1 ORAI calcium release-activated calcium modulator 1 SEQID NO: 9825

ORM1 Orosomucoid 1 SEQID NO: 9826

ORM2 Orosomucoid 2 SEQID NO: 9827

ORMDL2 ORMDL sphingolipid biosynthesis regulator 2 SEQID NOS: 9828- 9831

OS9 Osteosarcoma amplified 9, endoplasmic reticulum SEQID NOS: 9832 -9846 lectin

OSCAR Osteoclast associated, immunoglobulin-like receptor SEQID NOS: 9847-9857

OSM Oncostatin M SEQID NOS: 9858-9860

OSMR Oncostatin M receptor SEQID NOS: 9861 -9865

OSTN Osteocrin SEQID NOS: 9866-9867

OTOA Otoancorin SEQID NOS: 9868-9873

OTOG Otogelin SEQID NOS: 9874-9876

OTOGL Otogelin-like SEQID NOS: 9877-9883

OTOL1 Otolin 1 SEQID NO: 9884

OTOR Otoraplin SEQID NO: 9885

OTOS Otospiralin SEQID NOS: 9886-9887

OVCH1 Ovochymase 1 SEQID NOS: 9888-9890

OVCH2 Ovochymase 2 (gene/pseudogene) SEQID NOS: 9891 -9892

OVGP1 Oviductal glycoprotein 1, 120kDa SEQID NO: 9893

OXCT1 3-oxoacid CoA transferase 1 SEQID NOS: 9894-9897

OXCT2 3-oxoacid CoA transferase 2 SEQID NO: 9898

OXNAD1 Oxidoreductase NAD-binding domain containing 1 SEQID NOS: 9899-9905

OXT Oxytocin/neurophysin 1 prepropeptide SEQID NO: 9906

P3H1 Prolyl 3-hydroxylase 1 SEQID NOS: 9907-9911

P3H2 Prolyl 3-hydroxylase 2 SEQID NOS: 9912 -9915

P3H3 Prolyl 3-hydroxylase 3 SEQID NO: 9916

P3H4 Prolyl 3-hydroxylase family member 4 (non- SEQID NOS: 9917-9921 enzymatic)

P4HA1 Prolyl 4-hydroxylase, alpha polypeptide 1 SEQID NOS: 9922 -9926

P4HA2 Prolyl 4-hydroxylase, alpha polypeptide II SEQID NOS: 9927-9941 P4HA3 Prolyl 4-hydroxylase, alpha polypeptide III SEQID NOS: 9942-9946

P4HB Prolyl 4-hydroxylase, beta polypeptide SEQID NOS: 9947-9958

PAEP Progestagen-associated endometrial protein SEQID NOS: 9959-9967

PAM Peptidylglycine alpha-amidating monooxygenase SEQID NOS: 9968-9981

PAMR1 Peptidase domain containing associated with muscle SEQID NOS: 9982-9988 regeneration 1

PAPLN Papilin, proteoglycan-like sulfated glycoprotein SEQID NOS: 9989-9996

PAPPA Pregnancy-associated plasma protein A, pappalysin SEQID NO: 9997

1

PAPPA2 Pappalysin 2 SEQID NOS: 9998-9999

PARP15 Poly (ADP-ribose) polymerase family, member 15 SEQID NOS: 10000- 10003

PARVB Parvin, beta SEQID NOS: 10004- 10008

PATE1 Prostate and testis expressed 1 SEQID NOS: 10009- 10010

PATE2 Prostate and testis expressed 2 SEQID NOS: 10011- 10012

PATE3 Prostate and testis expressed 3 SEQID NO: 10013

PATE4 Prostate and testis expressed 4 SEQID NOS: 10014- 10015

PATL2 Protein associated with topoisomerase II homolog 2 SEQID NOS: 10016- 10021

(yeast)

PAX2 Paired box 2 SEQID NOS: 10022- 10027

PAX4 Paired box 4 SEQID NOS: 10028- 10034

PCCB Propionyl CoA carboxylase, beta polypeptide SEQID NOS: 10035- 10049

PCDH1 Protocadherin 1 SEQID NOS: 10050- 10055

PCDH12 Protocadherin 12 SEQID NOS: 10056- 10057

PCDH15 Protocadherin-related 15 SEQID NOS: 10058- 10091

PCDHA1 Protocadherin alpha 1 SEQID NOS: 10092- 10094

PCDHA10 Protocadherin alpha 10 SEQID NOS: 10095- 10097

PCDHA11 Protocadherin alpha 11 SEQID NOS: 10098- 10100

PCDHA6 Protocadherin alpha 6 SEQID NOS: 10101-10103

PCDHB12 Protocadherin beta 12 SEQID NOS: 10104-10106

PCDHGA11 Protocadherin gamma subfamily A, 11 SEQID NOS: 10107-10109

PCF11 PCF11 cleavage and polyadenylation factor subunit SEQID NOS: 10110-10114

PCOLCE Procollagen C-endopeptidase enhancer SEQID NO: 10115

PCOLCE2 Procollagen C-endopeptidase enhancer 2 SEQID NOS: 10116-10119

PCSK1 Proprotein convertase subtilisin/kexin type 1 SEQID NOS: 10120-10122

PCSK1N Proprotein convertase subtilisin/kexin type 1 SEQID NO: 10123

inhibitor

PCSK2 Proprotein convertase subtilisin/kexin type 2 SEQID NOS: 10124-10126

PCSK4 Proprotein convertase subtilisin/kexin type 4 SEQID NOS: 10127-10129

PCSK5 Proprotein convertase subtilisin/kexin type 5 SEQID NOS: 10130-10134

PCSK9 Proprotein convertase subtilisin/kexin type 9 SEQID NO: 10135

PCYOX1 Prenylcysteine oxidase 1 SEQID NOS: 10136-10140

PCYOX1L Prenylcysteine oxidase 1 like SEQID NOS: 10141-10145

PDE11A Phosphodiesterase 11A SEQID NOS: 10146-10151

PDE2A Phosphodiesterase 2A, cGMP-stimulated SEQID NOS: 10152-10173

PDE7A Phosphodiesterase 7A SEQID NOS: 10174-10177

PDF Peptide deformylase (mitochondrial) SEQID NO: 10178

PDGFA Platelet-derived growth factor alpha polypeptide SEQID NOS: 10179-10182 PDGFB Platelet-derived growth factor beta polypeptide SEQID NOS: 10183 - 10186

PDGFC Platelet derived growth factor C SEQID NOS: 10187- 10190

PDGFD Platelet derived growth factor D SEQID NOS: 10191 - 10193

PDGFRA Platelet-derived growth factor receptor, alpha SEQID NOS: 10194- 10200 polypeptide

PDGFRB Platelet-derived growth factor receptor, beta SEQID NOS: 10201 - 10204 polypeptide

PDGFRL Platelet-derived growth factor receptor-like SEQID NOS: 10205 - 10206

PDHA1 Pyruvate dehydrogenase (lipoamide) alpha 1 SEQID NOS: 10207- 10215

PDIA2 Protein disulfide isomerase family A, member 2 SEQID NOS: 10216- 10219

PDIA3 Protein disulfide isomerase family A, member 3 SEQID NOS: 10220- 10223

PDIA4 Protein disulfide isomerase family A, member 4 SEQID NOS: 10224- 10225

PDIA5 Protein disulfide isomerase family A, member 5 SEQID NOS: 10226- 10229

PDIA6 Protein disulfide isomerase family A, member 6 SEQID NOS: 10230- 10236

PDILT Protein disulfide isomerase-like, testis expressed SEQID NOS: 10237- 10238

PDYN Prodynorphin SEQID NOS: 10239- 10241

PDZD8 PDZ domain containing 8 SEQID NO: 10242

PDZRN4 PDZ domain containing ring finger 4 SEQID NOS: 10243 - 10245

PEAR1 Platelet endothelial aggregation receptor 1 SEQID NOS: 10246- 10249

PEBP4 Phosphatidylethanolamine-binding protein 4 SEQID NOS: 10250- 10251

PECAM1 Platelet/endothelial cell adhesion molecule 1 SEQID NOS: 10252 - 10255

PENK Proenkephalin SEQID NOS: 10256- 10261

PET117 PET117 homolog SEQID NO: 10262

PF4 Platelet factor 4 SEQID NO: 10263

PF4V1 Platelet factor 4 variant 1 SEQID NO: 10264

PFKP Phosphofructokinase, platelet SEQID NOS: 10265 - 10273

PFN1 Profilin 1 SEQID NOS: 10274- 10276

PGA3 Pepsinogen 3, group 1 (pepsinogen A) SEQID NOS: 10277- 10280

PGA4 Pepsinogen 4, group 1 (pepsinogen A) SEQID NOS: 10281 - 10283

PGA5 Pepsinogen 5, group 1 (pepsinogen A) SEQID NOS: 10284- 10286

PGAM5 PGAM family member 5, serine/threonine protein SEQID NOS: 10287- 10290 phosphatase, mitochondrial

PGAP3 Post-GPI attachment to proteins 3 SEQID NOS: 10291 - 10298

PGC Progastricsin (pepsinogen C) SEQID NOS: 10299- 10302

PGF Placental growth factor SEQID NOS: 10303 - 10306

PGLYRP1 Peptidoglycan recognition protein 1 SEQID NO: 10307

PGLYRP2 Peptidoglycan recognition protein 2 SEQID NOS: 10308- 10311

PGLYRP3 Peptidoglycan recognition protein 3 SEQID NO: 10312

PGLYRP4 Peptidoglycan recognition protein 4 SEQID NOS: 10313 - 10314

PHACTR1 Phosphatase and actin regulator 1 SEQID NOS: 10315 - 10321

PHB Prohibitin SEQID NOS: 10322 - 10330

PI15 Peptidase inhibitor 15 SEQID NOS: 10331 - 10332

PI3 Peptidase inhibitor 3, skin-derived SEQID NO: 10333

PIANP PILR alpha associated neural protein SEQID NOS: 10334- 10339

PIGK Phosphatidylinositol glycan anchor biosynthesis, SEQID NOS: 10340- 10343 class K PIGL Phosphatidylinositol glycan anchor biosynthesis, SEQID NOS: 10344- 10351 class L

PIGT Phosphatidylinositol glycan anchor biosynthesis, SEQID NOS: 10352 - 10406 class T

PIGZ Phosphatidylinositol glycan anchor biosynthesis, SEQID NOS: 10407-10409 class Z

PIK3AP1 Phosphoinositide-3-kinase adaptor protein 1 SEQID NOS: 10410-10412

PIK3IP1 Phosphoinositide-3-kinase interacting protein 1 SEQID NOS: 10413-10416

PILRA Paired immunoglobin-like type 2 receptor alpha SEQID NOS: 10417-10421

PILRB Paired immunoglobin-like type 2 receptor beta SEQID NOS: 10422- 10433

PINLYP Phospholipase A2 inhibitor and LY6/PLAUR domain SEQID NOS: 10434-10438 containing

PIP Prolactin-induced protein SEQID NO: 10439

PIWIL4 Piwi-like RNA-mediated gene silencing 4 SEQID NOS: 10440-10444

PKDCC Protein kinase domain containing, cytoplasmic SEQID NOS: 10445-10446

PKHD1 Polycystic kidney and hepatic disease 1 (autosomal SEQID NOS: 10447-10448 recessive)

PLA1A Phospholipase Al member A SEQID NOS: 10449-10453

PLA2G10 Phospholipase A2, group X SEQID NOS: 10454-10455

PLA2G12A Phospholipase A2, group XI IA SEQID NOS: 10456- 10458

PLA2G12B Phospholipase A2, group XI IB SEQID NO: 10459

PLA2G15 Phospholipase A2, group XV SEQID NOS: 10460-10467

PLA2G1B Phospholipase A2, group IB (pancreas) SEQID NOS: 10468-10470

PLA2G2A Phospholipase A2, group IIA (platelets, synovial SEQID NOS: 10471 - 10472 fluid)

PLA2G2C Phospholipase A2, group IIC SEQID NOS: 10473- 10474

PLA2G2D Phospholipase A2, group IID SEQID NOS: 10475- 10476

PLA2G2E Phospholipase A2, group HE SEQID NO: 10477

PLA2G3 Phospholipase A2, group III SEQID NO: 10478

PLA2G5 Phospholipase A2, group V SEQID NO: 10479

PLA2G7 Phospholipase A2, group VII (platelet-activating SEQID NOS: 10480-10481 factor acetylhydrolase, plasma)

PLA2R1 Phospholipase A2 receptor 1, 180kDa SEQID NOS: 10482- 10483

PLAC1 Placenta-specific 1 SEQID NO: 10484

PLAC9 Placenta-specific 9 SEQID NOS: 10485- 10487

PLAT Plasminogen activator, tissue SEQID NOS: 10488-10496

PLAU Plasminogen activator, urokinase SEQID NOS: 10497-10499

PLAUR Plasminogen activator, urokinase receptor SEQID NOS: 10500- 10511

PLBD1 Phospholipase B domain containing 1 SEQID NOS: 10512 - 10514

PLBD2 Phospholipase B domain containing 2 SEQID NOS: 10515 - 10517

PLG Plasminogen SEQID NOS: 10518- 10520

PLGLB1 Plasminogen-like Bl SEQID NOS: 10521 - 10524

PLGLB2 Plasminogen-like B2 SEQID NOS: 10525 - 10526

PL0D1 Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 1 SEQID NOS: 10527- 10529

PL0D2 Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 2 SEQID NOS: 10530- 10535

PL0D3 Procollagen-lysine, 2-oxoglutarate 5-dioxygenase 3 SEQID NOS: 10536- 10542

PLTP Phospholipid transfer protein SEQID NOS: 10543- 10547 PLXNA4 Plexin A4 SEQID NOS: 10548- 10551

PLXNB2 Plexin B2 SEQID NOS: 10552 - 10560

PM20D1 Peptidase M20 domain containing 1 SEQID NO: 10561

PMCH Pro-melanin-concentrating hormone SEQID NO: 10562

PMEL Premelanosome protein SEQID NOS: 10563- 10574

PMEPA1 Prostate transmembrane protein, androgen induced SEQID NOS: 10575 - 10581

1

PNLIP Pancreatic lipase SEQID NO: 10582

PNLIPRPl Pancreatic lipase-related protein 1 SEQID NOS: 10583- 10591

PNLIPRP3 Pancreatic lipase-related protein 3 SEQID NO: 10592

PNOC Prepronociceptin SEQID NOS: 10593- 10595

PNP Purine nucleoside phosphorylase SEQID NOS: 10596- 10599

PNPLA4 Patatin-like phospholipase domain containing 4 SEQID NOS: 10600- 10603

PODNL1 Podocan-like 1 SEQID NOS: 10604- 10615

POFUT1 Protein O-fucosyltransferase 1 SEQID NOS: 10616- 10617

POFUT2 Protein O-fucosyltransferase 2 SEQID NOS: 10618- 10623

POGLUT1 Protein O-glucosyltransferase 1 SEQID NOS: 10624- 10628

POLL Polymerase (DNA directed), lambda SEQID NOS: 10629- 10641

POMC Proopiomelanocortin SEQID NOS: 10642- 10646

POMGNT2 Protein O-linked mannose N- SEQID NOS: 10647-10648 acetylglucosaminyltransferase 2 (beta 1,4-)

PON1 Paraoxonase 1 SEQID NOS: 10649- 10650

PON2 Paraoxonase 2 SEQID NOS: 10651- 10663

PON3 Paraoxonase 3 SEQID NOS: 10664- 10669

POSTN Periostin, osteoblast specific factor SEQID NOS: 10670- 10675

PPBP Pro-platelet basic protein (chemokine (C-X-C motif) SEQID NO: 10676

ligand 7)

PPIB Peptidylprolyl isomerase B (cyclophilin B) SEQID NO: 10677

PPIC Peptidylprolyl isomerase C (cyclophilin C) SEQID NO: 10678

PPOX Protoporphyrinogen oxidase SEQID NOS: 10679- 10689

PPP1CA Protein phosphatase 1, catalytic subunit, alpha SEQID NOS: 10690- 10695 isozyme

PPT1 Palmitoyl-protein thioesterase 1 SEQID NOS: 10696- 10712

PPT2 Palmitoyl-protein thioesterase 2 SEQID NOS: 10713- 10720

PPY Pancreatic polypeptide SEQID NOS: 10721- 10725

PRAC2 Prostate cancer susceptibility candidate 2 SEQID NOS: 10726- 10727

PRADC1 Protease-associated domain containing 1 SEQID NO: 10728

PRAP1 Proline-rich acidic protein 1 SEQID NOS: 10729- 10730

PRB1 Proline-rich protein BstNI subfamily 1 SEQID NOS: 10731- 10734

PRB2 Proline-rich protein BstNI subfamily 2 SEQID NOS: 10735- 10736

PRB3 Proline-rich protein BstNI subfamily 3 SEQID NOS: 10737- 10738

PRB4 Proline-rich protein BstNI subfamily 4 SEQID NOS: 10739- 10742

PRCD Progressive rod-cone degeneration SEQID NOS: 10743- 10744

PRCP Prolylcarboxypeptidase (angiotensinase C) SEQID NOS: 10745- 10756

PRDM12 PR domain containing 12 SEQID NO: 10757

PRDX4 Peroxiredoxin 4 SEQID NOS: 10758- 10761

PRELP Proline/arginine-rich end leucine-rich repeat protein SEQID NO: 10762 PRF1 Perforin 1 (pore forming protein) SEQID NOS: 10763 - 10765

PRG2 Proteoglycan 2, bone marrow (natural killer cell SEQID NOS: 10766- 10768 activator, eosinophil granule major basic protein)

PRG3 Proteoglycan 3 SEQID NO: 10769

PRG4 Proteoglycan 4 SEQID NOS: 10770- 10775

PRH1 Proline-rich protein Haelll subfamily 1 SEQID NOS: 10776- 10778

PRH2 Proline-rich protein Haelll subfamily 2 SEQID NOS: 10779- 10780

PRKAG1 Protein kinase, AMP-activated, gamma 1 non- SEQID NOS: 10781 - 10795 catalytic subunit

PRKCSH Protein kinase C substrate 80K-H SEQID NOS: 10796- 10805

PRKD1 Protein kinase Dl SEQID NOS: 10806- 10811

PRL Prolactin SEQID NOS: 10812- 10814

PRLH Prolactin releasing hormone SEQID NO: 10815

PRLR Prolactin receptor SEQID NOS: 10816- 10834

PRNP Prion protein SEQID NOS: 10835- 10838

PRNT Prion protein (testis specific) SEQID NO: 10839

PROC Protein C (inactivator of coagulation factors Va and SEQID NOS: 10840-10847

Villa)

PR0K1 Prokineticin 1 SEQID NO: 10848

PROK2 Prokineticin 2 SEQID NOS: 10849- 10850

PROM1 Prominin 1 SEQID NOS: 10851- 10862

PROS1 Protein S (alpha) SEQID NOS: 10863- 10866

PROZ Protein Z, vitamin K-dependent plasma glycoprotein SEQID NOS: 10867- 10868

PRR27 Proline rich 27 SEQID NOS: 10869- 10872

PRR4 Proline rich 4 (lacrimal) SEQID NOS: 10873- 10875

PRRG2 Proline rich Gla (G-carboxyglutamic acid) 2 SEQID NOS: 10876- 10878

PRRT3 Proline-rich transmembrane protein 3 SEQID NOS: 10879- 10881

PRRT4 Proline-rich transmembrane protein 4 SEQID NOS: 10882- 10888

PRSS1 Protease, serine, 1 (trypsin 1) SEQID NOS: 10889- 10892

PRSS12 Protease, serine, 12 (neurotrypsin, motopsin) SEQID NO: 10893

PRSS16 Protease, serine, 16 (thymus) SEQID NOS: 10894- 10901

PRSS2 Protease, serine, 2 (trypsin 2) SEQID NOS: 10902- 10905

PRSS21 Protease, serine, 21 (testisin) SEQID NOS: 10906- 10911

PRSS22 Protease, serine, 22 SEQID NOS: 10912- 10914

PRSS23 Protease, serine, 23 SEQID NOS: 10915- 10918

PRSS27 Protease, serine 27 SEQID NOS: 10919- 10921

PRSS3 Protease, serine, 3 SEQID NOS: 10922- 10926

PRSS33 Protease, serine, 33 SEQID NOS: 10927- 10930

PRSS35 Protease, serine, 35 SEQID NO: 10931

PRSS36 Protease, serine, 36 SEQID NOS: 10932- 10935

PRSS37 Protease, serine, 37 SEQID NOS: 10936- 10939

PRSS38 Protease, serine, 38 SEQID NO: 10940

PRSS42 Protease, serine, 42 SEQID NOS: 10941-10942

PRSS48 Protease, serine, 48 SEQID NOS: 10943- 10944

PRSS50 Protease, serine, 50 SEQID NO: 10945

PRSS53 Protease, serine, 53 SEQID NO: 10946

PRSS54 Protease, serine, 54 SEQID NOS: 10947- 10951 PRSS55 Protease, serine, 55 SEQID NOS: 10952 - 10954

PRSS56 Protease, serine, 56 SEQIDNOS: 10955- 10956

PRSS57 Protease, serine, 57 SEQID NOS: 10957- 10958

PRSS58 Protease, serine, 58 SEQID NOS: 10959- 10960

PRSS8 Protease, serine, 8 SEQIDNOS: 10961- 10964

PRTG Protogenin SEQIDNOS: 10965- 10968

PRTN3 Proteinase 3 SEQID NOS: 10969- 10970

PSAP Prosaposin SEQIDNOS: 10971- 10974

PSAPL1 Prosaposin-like 1 (gene/pseudogene) SEQID NO: 10975

PSG1 Pregnancy specific beta-l-glycoprotein 1 SEQID NOS: 10976- 10983

PSG11 Pregnancy specific beta-l-glycoprotein 11 SEQID NOS: 10984- 10988

PSG2 Pregnancy specific beta-l-glycoprotein 2 SEQID NOS: 10989- 10990

PSG3 Pregnancy specific beta-l-glycoprotein 3 SEQIDNOS: 10991- 10994

PSG4 Pregnancy specific beta-l-glycoprotein 4 SEQIDNOS: 10995- 11006

PSG5 Pregnancy specific beta-l-glycoprotein 5 SEQID NOS: 11007- 11012

PSG6 Pregnancy specific beta-l-glycoprotein 6 SEQIDNOS: 11013-11018

PSG7 Pregnancy specific beta-l-glycoprotein 7 SEQID NOS: 11019-11021

(gene/pseudogene)

PSG8 Pregnancy specific beta-l-glycoprotein 8 SEQIDNOS: 11022 - 11026

PSG9 Pregnancy specific beta-l-glycoprotein 9 SEQID NOS: 11027- 11034

PSMD1 Proteasome 26S subunit, non-ATPase 1 SEQIDNOS: 11035- 11042

PS0RS1C2 Psoriasis susceptibility 1 candidate 2 SEQID NO: 11043

PSPN Persephin SEQID NOS: 11044-11045

PTGDS Prostaglandin D2 synthase 21kDa (brain) SEQID NOS: 11046- 11050

PTGIR Prostaglandin 12 (prostacyclin) receptor (IP) SEQIDNOS: 11051- 11055

PTGS1 Prostaglandin-endoperoxide synthase 1 SEQID NOS: 11056- 11064

(prostaglandin G/H synthase and cyclooxygenase)

PTGS2 Prostaglandin-endoperoxide synthase 2 SEQID NOS: 11065 - 11066

(prostaglandin G/H synthase and cyclooxygenase)

PTH Parathyroid hormone SEQID NOS: 11067- 11068

PTH2 Parathyroid hormone 2 SEQID NO: 11069

PTHLH Parathyroid hormone-like hormone SEQID NOS: 11070- 11078

PTK7 Protein tyrosine kinase 7 (inactive) SEQID NOS: 11079- 11094

PTN Pleiotrophin SEQIDNOS: 11095- 11096

PTPRA Protein tyrosine phosphatase, receptor type, A SEQID NOS: 11097- 11104

PTPRB Protein tyrosine phosphatase, receptor type, B SEQIDNOS: 11105-11112

PTPRC Protein tyrosine phosphatase, receptor type, C SEQIDNOS: 11113-11123

PTPRCAP Protein tyrosine phosphatase, receptor type, C- SEQID NO: 11124

associated protein

PTPRD Protein tyrosine phosphatase, receptor type, D SEQIDNOS: 11125-11136

PTPRF Protein tyrosine phosphatase, receptor type, F SEQID NOS: 11137-11144

PTPRJ Protein tyrosine phosphatase, receptor type, J SEQIDNOS: 11145-11150

PTPRO Protein tyrosine phosphatase, receptor type, 0 SEQIDNOS: 11151-11159

PTPRS Protein tyrosine phosphatase, receptor type, S SEQID NOS: 11160-11167

PTTG1IP Pituitary tumor-transforming 1 interacting protein SEQID NOS: 11168-11171

PTX3 Pentraxin 3, long SEQID NO: 11172

PTX4 Pentraxin 4, long SEQIDNOS: 11173-11175 PVR Poliovirus receptor SEQID NOS: 11176-11181

PXDN Peroxidasin SEQIDNOS: 11182-11186

PXDNL Peroxidasin-like SEQID NOS: 11187-11189

PXYLP1 2-phosphoxylose phosphatase 1 SEQID NOS: 11190- 11202

PYY Peptide YY SEQIDNOS: 11203 - 11204

PZP Pregnancy-zone protein SEQIDNOS: 11205 - 11206

QPCT Glutaminyl-peptide cyclotransferase SEQID NOS: 11207- 11209

QPRT Quinolinate phosphoribosyltransferase SEQID NOS: 11210-11211

QRFP Pyroglutamylated RFamide peptide SEQIDNOS: 11212-11213

QS0X1 Quiescin Q6 sulfhydryl oxidase 1 SEQID NOS: 11214-11217

R3HDML R3H domain containing-like SEQID NO: 11218

RAB26 RAB26, member RAS oncogene family SEQID NOS: 11219-11222

RAB36 RAB36, member RAS oncogene family SEQIDNOS: 11223 - 11225

RAB9B RAB9B, member RAS oncogene family SEQID NO: 11226

RAET1E Retinoic acid early transcript IE SEQID NOS: 11227- 11232

RAET1G Retinoic acid early transcript 1G SEQIDNOS: 11233 - 11235

RAMP2 Receptor (G protein-coupled) activity modifying SEQID NOS: 11236- 11240 protein 2

RAPGEF5 Rap guanine nucleotide exchange factor (GEF) 5 SEQIDNOS: 11241-11247

RARRES1 Retinoic acid receptor responder (tazarotene SEQID NOS: 11248- 11249 induced) 1

RARRES2 Retinoic acid receptor responder (tazarotene SEQID NOS: 11250- 11253 induced) 2

RASA 2 RAS p21 protein activator 2 SEQID NOS: 11254- 11256

RBM3 RNA binding motif (RNP1, RRM) protein 3 SEQID NOS: 11257- 11259

RBP3 Retinol binding protein 3, interstitial SEQID NO: 11260

RBP4 Retinol binding protein 4, plasma SEQIDNOS: 11261- 11264

RCN1 Reticulocalbin 1, EF-hand calcium binding domain SEQIDNOS: 11265 - 11268

RCN2 Reticulocalbin 2, EF-hand calcium binding domain SEQID NOS: 11269- 11272

RCN3 Reticulocalbin 3, EF-hand calcium binding domain SEQIDNOS: 11273 - 11276

RC0R1 REST corepressor 1 SEQID NOS: 11277- 11278

RDH11 Retinol dehydrogenase 11 (all-trans/9-cis/ll-cis) SEQID NOS: 11279- 11286

RDH12 Retinol dehydrogenase 12 (all-trans/9-cis/ll-cis) SEQID NOS: 11287- 11288

RDH13 Retinol dehydrogenase 13 (all-trans/9-cis) SEQID NOS: 11289- 11297

RDH5 Retinol dehydrogenase 5 (ll-cis/9-cis) SEQID NOS: 11298- 11302

RDH8 Retinol dehydrogenase 8 (all-trans) SEQIDNOS: 11303 - 11304

REG1A Regenerating islet-derived 1 alpha SEQID NO: 11305

REG1B Regenerating islet-derived 1 beta SEQID NOS: 11306- 11307

REG3A Regenerating islet-derived 3 alpha SEQID NOS: 11308- 11310

REG3G Regenerating islet-derived 3 gamma SEQIDNOS: 11311-11313

REG4 Regenerating islet-derived family, member 4 SEQID NOS: 11314-11317

RELN Reelin SEQID NOS: 11318-11321

RELT RELT tumor necrosis factor receptor SEQIDNOS: 11322 - 11325

REN Renin SEQID NOS: 11326- 11327

REPIN1 Replication initiator 1 SEQID NOS: 11328- 11341

REPS2 RALBP1 associated Eps domain containing 2 SEQIDNOS: 11342- 11343

RET Ret proto-oncogene SEQID NOS: 11344-11349 RETN Resistin SEQID NOS: 11350- 11352

RETNLB Resistin like beta SEQID NO: 11353

RETSAT Retinol saturase (all-trans-retinol 13,14-reductase) SEQID NOS: 11354- 11358

RFNG RFNG O-fucosylpeptide 3-beta-N- SEQID NOS: 11359- 11361 acetylglucosaminyltransferase

RGCC Regulator of cell cycle SEQID NO: 11362

RGL4 Ral guanine nucleotide dissociation stimulator-like 4 SEQID NOS: 11363 - 11369

RGMA Repulsive guidance molecule family member a SEQID NOS: 11370- 11379

RGMB Repulsive guidance molecule family member b SEQID NOS: 11380- 11381

RHOQ Ras homolog family member Q SEQID NOS: 11382 - 11386

RIC3 RIC3 acetylcholine receptor chaperone SEQID NOS: 11387- 11394

HRSP12 Heat-responsive protein 12 SEQID NOS: 11395 - 11398

RIMS1 Regulating synaptic membrane exocytosis 1 SEQID NOS: 11399- 11414

RIPPLY1 Ripply transcriptional repressor 1 SEQID NOS: 11415-11416

RLN1 Relaxin 1 SEQID NO: 11417

RLN2 Relaxin 2 SEQID NOS: 11418-11419

RLN3 Relaxin 3 SEQID NOS: 11420- 11421

RMDN1 Regulator of microtubule dynamics 1 SEQID NOS: 11422- 11435

RNASE1 Ribonuclease, RNase A family, 1 (pancreatic) SEQID NOS: 11436-11440

RNASE10 Ribonuclease, RNase A family, 10 (non-active) SEQID NOS: 11441-11442

RNASE11 Ribonuclease, RNase A family, 11 (non-active) SEQID NOS: 11443-11453

RNASE12 Ribonuclease, RNase A family, 12 (non-active) SEQID NO: 11454

RNASE13 Ribonuclease, RNase A family, 13 (non-active) SEQID NO: 11455

RNASE2 Ribonuclease, RNase A family, 2 (liver, eosinophil- SEQID NO: 11456

derived neurotoxin)

RNASE3 Ribonuclease, RNase A family, 3 SEQID NO: 11457

RNASE4 Ribonuclease, RNase A family, 4 SEQID NOS: 11458-11460

RNASE6 Ribonuclease, RNase A family, k6 SEQID NO: 11461

RNASE7 Ribonuclease, RNase A family, 7 SEQID NOS: 11462-11463

RNASE8 Ribonuclease, RNase A family, 8 SEQID NO: 11464

RNASE9 Ribonuclease, RNase A family, 9 (non-active) SEQID NOS: 11465-11475

RNASEH1 Ribonuclease HI SEQID NOS: 11476-11478

RNASET2 Ribonuclease T2 SEQID NOS: 11479-11486

RNF146 Ring finger protein 146 SEQID NOS: 11487-11498

RNF148 Ring finger protein 148 SEQID NOS: 11499- 11500

RNF150 Ring finger protein 150 SEQID NOS: 11501- 11505

RNF167 Ring finger protein 167 SEQID NOS: 11506- 11516

RNF220 Ring finger protein 220 SEQID NOS: 11517-11523

RNF34 Ring finger protein 34, E3 ubiquitin protein ligase SEQID NOS: 11524- 11531

RNLS Renalase, FAD-dependent amine oxidase SEQID NOS: 11532 - 11534

RNPEP Arginyl aminopeptidase (aminopeptidase B) SEQID NOS: 11535- 11540

R0R1 Receptor tyrosine kinase-like orphan receptor 1 SEQID NOS: 11541-11543

RPL3 Ribosomal protein L3 SEQID NOS: 11544-11549

RPLP2 Ribosomal protein, large, P2 SEQID NOS: 11550- 11552

RPN2 Ribophorin II SEQID NOS: 11553 - 11559

RPS27L Ribosomal protein S27-like SEQID NOS: 11560- 11565

RSI Retinoschisin 1 SEQID NO: 11566 RSF1 Remodeling and spacing factor 1 SEQID NOS: 11567- 11573

RSPOl R-spondin 1 SEQID NOS: 11574- 11577

RSP02 R-spondin 2 SEQID NOS: 11578- 11585

RSP03 R-spondin 3 SEQID NOS: 11586- 11587

RSP04 R-spondin 4 SEQID NOS: 11588- 11589

RSPRY1 Ring finger and SPRY domain containing 1 SEQID NOS: 11590- 11596

RTBDN Retbindin SEQID NOS: 11597- 11609

RTN4RL1 Reticulon 4 receptor-like 1 SEQID NO: 11610

RTN4RL2 Reticulon 4 receptor-like 2 SEQID NOS: 11611 - 11613

SAA1 Serum amyloid Al SEQID NOS: 11614- 11616

SAA2 Serum amyloid A2 SEQID NOS: 11617- 11622

SAA4 Serum amyloid A4, constitutive SEQID NO: 11623

SAP30 Sin3A-associated protein, 30kDa SEQID NO: 11624

SAR1A Secretion associated, Ras related GTPase 1A SEQID NOS: 11625 - 11631

SARAF Store-operated calcium entry-associated regulatory SEQID NOS: 11632 - 11642 factor

SARM1 Sterile alpha and TIR motif containing 1 SEQID NOS: 11643 - 11646

SATB1 SATB homeobox 1 SEQID NOS: 11647- 11659

SAX 02 Stabilizer of axonemal microtubules 2 SEQID NOS: 11660- 11664

SBSN Suprabasin SEQID NOS: 11665 - 11667

SBSPON Somatomedin B and thrombospondin, type 1 SEQID NO: 11668

domain containing

SCARF1 Scavenger receptor class F, member 1 SEQID NOS: 11669- 11673

SCG2 Secretogranin II SEQID NOS: 11674- 11676

SCG3 Secretogranin III SEQID NOS: 11677- 11679

SCG5 Secretogranin V SEQID NOS: 11680- 11684

SCGB1A1 Secretoglobin, family 1A, member 1 (uteroglobin) SEQID NOS: 11685- 11686

SCGB1C1 Secretoglobin, family 1C, member 1 SEQID NO: 11687

SCGB1C2 Secretoglobin, family 1C, member 2 SEQID NO: 11688

SCGB1D1 Secretoglobin, family ID, member 1 SEQID NO: 11689

SCGB1D2 Secretoglobin, family ID, member 2 SEQID NO: 11690

SCGB1D4 Secretoglobin, family ID, member 4 SEQID NO: 11691

SCGB2A1 Secretoglobin, family 2A, member 1 SEQID NO: 11692

SCGB2A2 Secretoglobin, family 2A, member 2 SEQID NOS: 11693 - 11694

SCGB2B2 Secretoglobin, family 2B, member 2 SEQID NOS: 11695 - 11696

SCGB3A1 Secretoglobin, family 3A, member 1 SEQID NO: 11697

SCGB3A2 Secretoglobin, family 3A, member 2 SEQID NOS: 11698- 11699

SCN1B Sodium channel, voltage gated, type 1 beta subunit SEQID NOS: 11700- 11705

SCN3B Sodium channel, voltage gated, type III beta subunit SEQID NOS: 11706- 11710

SCPEP1 Serine carboxypeptidase 1 SEQID NOS: 11711 - 11718

SCRG1 Stimulator of chondrogenesis 1 SEQID NOS: 11719- 11720

SCT Secretin SEQID NO: 11721

SCUBE1 Signal peptide, CUB domain, EGF-like 1 SEQID NOS: 11722 - 11725

SCUBE2 Signal peptide, CUB domain, EGF-like 2 SEQID NOS: 11726- 11732

SCUBE3 Signal peptide, CUB domain, EGF-like 3 SEQID NO: 11733

SDC1 Syndecan 1 SEQID NOS: 11734- 11738

SDF2 Stromal cell-derived factor 2 SEQID NOS: 11739- 11741 SDF2L1 Stromal cell-derived factor 2-like 1 SEQID NO: 11742

SDF4 Stromal cell derived factor 4 SEQIDNOS: 11743-11746

SDHAF2 Succinate dehydrogenase complex assembly factor 2 SEQID NOS: 11747- 11754

SDHAF4 Succinate dehydrogenase complex assembly factor 4 SEQID NO: 11755

SDHB Succinate dehydrogenase complex, subunit B, iron SEQID NOS: 11756- 11758 sulfur (Ip)

SDHD Succinate dehydrogenase complex, subunit D, SEQID NOS: 11759- 11768 integral membrane protein

SEC14L3 SEC14-like lipid binding 3 SEQID NOS: 11769- 11775

SEC16A SEC16 homolog A, endoplasmic reticulum export SEQID NOS: 11776- 11782 factor

SEC16B SEC16 homolog B, endoplasmic reticulum export SEQIDNOS: 11783- 11786 factor

SEC22C SEC22 homolog C, vesicle trafficking protein SEQID NOS: 11787- 11799

SEC31A SEC31 homolog A, COPII coat complex component SEQID NOS: 11800- 11829

SECISBP2 SECIS binding protein 2 SEQID NOS: 11830- 11834

SECTM1 Secreted and transmembrane 1 SEQIDNOS: 11835- 11842

SEL1L Sel-1 suppressor of lin-12-like (C. elegans) SEQIDNOS: 11843-11845

SEPT15 15 kDa selenoprotein SEQID NOS: 11846- 11852

SELM Selenoprotein M SEQIDNOS: 11853 - 11855

SEPN1 Selenoprotein N, 1 SEQID NOS: 11856- 11859

SELO Selenoprotein 0 SEQID NOS: 11860- 11861

SEPP1 Selenoprotein P, plasma, 1 SEQIDNOS: 11862- 11867

SEMA3A Sema domain, immunoglobulin domain (Ig), short SEQID NOS: 11868- 11872 basic domain, secreted, (semaphorin) 3A

SEMA3B Sema domain, immunoglobulin domain (Ig), short SEQID NOS: 11873 - 11879 basic domain, secreted, (semaphorin) 3B

SEMA3C Sema domain, immunoglobulin domain (Ig), short SEQID NOS: 11880- 11884 basic domain, secreted, (semaphorin) 3C

SEMA3E Sema domain, immunoglobulin domain (Ig), short SEQID NOS: 11885 - 11889 basic domain, secreted, (semaphorin) 3E

SEMA3F Sema domain, immunoglobulin domain (Ig), short SEQID NOS: 11890- 11896 basic domain, secreted, (semaphorin) 3F

SEMA3G Sema domain, immunoglobulin domain (Ig), short SEQID NOS: 11897- 11899 basic domain, secreted, (semaphorin) 3G

SEMA4A Sema domain, immunoglobulin domain (Ig), SEQID NOS: 11900- 11908 transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4A

SEMA4B Sema domain, immunoglobulin domain (Ig), SEQID NOS: 11909- 11919 transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4B

SEMA4C Sema domain, immunoglobulin domain (Ig), SEQID NOS: 11920- 11922 transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4C

SEMA4D Sema domain, immunoglobulin domain (Ig), SEQID NOS: 11923 - 11936 transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4D SEMA4F Sema domain, immunoglobulin domain (Ig), SEQID NOS: 11937- 11945 transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4F

SEMA4G Sema domain, immunoglobulin domain (Ig), SEQID NOS: 11946- 11953 transmembrane domain (TM) and short cytoplasmic

domain, (semaphorin) 4G

SEMA5A Sema domain, seven thrombospondin repeats (type SEQID NOS: 11954- 11955

1 and type 1-like), transmembrane domain (TM) and

short cytoplasmic domain, (semaphorin) 5A

SEMA6A Sema domain, transmembrane domain (TM), and SEQID NOS: 11956- 11963 cytoplasmic domain, (semaphorin) 6A

SEMA6C Sema domain, transmembrane domain (TM), and SEQID NOS: 11964- 11969 cytoplasmic domain, (semaphorin) 6C

SEMA6D Sema domain, transmembrane domain (TM), and SEQID NOS: 11970- 11983 cytoplasmic domain, (semaphorin) 6D

SEMG1 Semenogelin 1 SEQID NO: 11984

SEMG2 Semenogelin II SEQID NO: 11985

SEPT9 Septin 9 SEQID NOS: 11986- 12022

SERPINA1 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12023 - 12039 antiproteinase, antitrypsin), member 1

SERPINA10 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12040- 12043 antiproteinase, antitrypsin), member 10

SERPINA11 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NO: 12044

antiproteinase, antitrypsin), member 11

SERPINA12 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12045 - 12046 antiproteinase, antitrypsin), member 12

SERPINA3 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12047- 12053 antiproteinase, antitrypsin), member 3

SERPINA4 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12054- 12056 antiproteinase, antitrypsin), member 4

SERPINA5 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12057- 12068 antiproteinase, antitrypsin), member 5

SERPINA6 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12069- 12071 antiproteinase, antitrypsin), member 6

SERPINA7 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12072 - 12073 antiproteinase, antitrypsin), member 7

SERPINA9 Serpin peptidase inhibitor, clade A (alpha-1 SEQID NOS: 12074- 12080 antiproteinase, antitrypsin), member 9

SERPINB2 Serpin peptidase inhibitor, clade B (ovalbumin), SEQID NOS: 12081 - 12085 member 2

SERPINC1 Serpin peptidase inhibitor, clade C (antithrombin), SEQID NOS: 12086- 12087 member 1

SERPIND1 Serpin peptidase inhibitor, clade D (heparin SEQID NOS: 12088- 12089 cofactor), member 1

SERPINE1 Serpin peptidase inhibitor, clade E (nexin, SEQID NO: 12090

plasminogen activator inhibitor type 1), member 1

SERPINE2 Serpin peptidase inhibitor, clade E (nexin, SEQID NOS: 12091 - 12097 plasminogen activator inhibitor type 1), member 2

SERPINE3 Serpin peptidase inhibitor, clade E (nexin, SEQID NOS: 12098- 12101 plasminogen activator inhibitor type 1), member 3 SERPINF1 Serpin peptidase inhibitor, clade F (alpha-2 SEQID NOS: 12102 - 12110 antiplasmin, pigment epithelium derived factor),

member 1

SERPINF2 Serpin peptidase inhibitor, clade F (alpha-2 SEQID NOS: 12111 - 12115 antiplasmin, pigment epithelium derived factor),

member 2

SERPING1 Serpin peptidase inhibitor, clade G (CI inhibitor), SEQID NOS: 12116- 12126 member 1

SERPINH1 Serpin peptidase inhibitor, clade H (heat shock SEQID NOS: 12127- 12141 protein 47), member 1, (collagen binding protein 1)

SERPINI1 Serpin peptidase inhibitor, clade 1 (neuroserpin), SEQID NOS: 12142 - 12146 member 1

SERPINI2 Serpin peptidase inhibitor, clade 1 (pancpin), SEQID NOS: 12147- 12153 member 2

SEZ6L2 Seizure related 6 homolog (mouse)-like 2 SEQID NOS: 12154- 12160

SFRP1 Secreted frizzled-related protein 1 SEQID NOS: 12161 - 12162

SFRP2 Secreted frizzled-related protein 2 SEQID NO: 12163

SFRP4 Secreted frizzled-related protein 4 SEQID NOS: 12164- 12165

SFRP5 Secreted frizzled-related protein 5 SEQID NO: 12166

SFTA2 Surfactant associated 2 SEQID NOS: 12167- 12168

SFTPA1 Surfactant protein Al SEQID NOS: 12169- 12173

SFTPA2 Surfactant protein A2 SEQID NOS: 12174- 12178

SFTPB Surfactant protein B SEQID NOS: 12179- 12183

SFTPD Surfactant protein D SEQID NOS: 12184- 12185

SFXN5 Sideroflexin 5 SEQID NOS: 12186- 12190

SGCA Sarcoglycan, alpha (50kDa dystrophin-associated SEQID NOS: 12191 - 12198 glycoprotein)

SGSH N-sulfoglucosamine sulfohydrolase SEQID NOS: 12199- 12207

SH3RF3 SH3 domain containing ring finger 3 SEQID NO: 12208

SHBG Sex hormone-binding globulin SEQID NOS: 12209- 12227

SHE Src homology 2 domain containing E SEQID NOS: 12228- 12230

SHH Sonic hedgehog SEQID NOS: 12231 - 12234

SHKBP1 SH3KBP1 binding protein 1 SEQID NOS: 12235 - 12250

SIAE Sialic acid acetylesterase SEQID NOS: 12251 - 12253

SIDT2 SID1 transmembrane family, member 2 SEQID NOS: 12254- 12263

SIGLEC10 Sialic acid binding Ig-like lectin 10 SEQID NOS: 12264- 12272

SIGLEC6 Sialic acid binding Ig-like lectin 6 SEQID NOS: 12273 - 12278

SIGLEC7 Sialic acid binding Ig-like lectin 7 SEQID NOS: 12279- 12283

SIGLECLl SIGLEC family like 1 SEQID NOS: 12284- 12289

SIGMAR1 Sigma non-opioid intracellular receptor 1 SEQID NOS: 12290- 12293

SIL1 SIL1 nucleotide exchange factor SEQID NOS: 12294- 12302

SIRPB1 Signal-regulatory protein beta 1 SEQID NOS: 12303 - 12315

SIRPD Signal-regulatory protein delta SEQID NOS: 12316- 12318

SLAMF1 Signaling lymphocytic activation molecule family SEQID NOS: 12319- 12321 member 1

SLAMF7 SLAM family member 7 SEQID NOS: 12322 - 12330

SLC10A3 Solute carrier family 10, member 3 SEQID NOS: 12331 - 12335 SLC15A3 Solute carrier family 15 (oligopeptide transporter), SEQ ID NOS: 12336 - 12341 member 3

SLC25A14 Solute carrier family 25 (mitochondrial carrier, SEQ ID NOS: 12342 - 12348 brain), member 14

SLC25A25 Solute carrier family 25 (mitochondrial carrier; SEQ ID NOS: 12349 - 12355 phosphate carrier), member 25

SLC2A5 Solute carrier family 2 (facilitated glucose/fructose SEQ ID NOS: 12356 - 12364 transporter), member 5

SLC35E3 Solute carrier family 35, member E3 SEQ ID NOS: 12365 - 12366

SLC39A10 Solute carrier family 39 (zinc transporter), member SEQ ID NOS: 12367 - 12373

10

SLC39A14 Solute carrier family 39 (zinc transporter), member SEQ ID NOS: 12374 - 12384

14

SLC39A4 Solute carrier family 39 (zinc transporter), member 4 SEQ ID NOS: 12385 - 12387

SLC39A5 Solute carrier family 39 (zinc transporter), member 5 SEQ ID NOS: 12388 - 12394

SLC3A1 Solute carrier family 3 (amino acid transporter heavy SEQ ID NOS: 12395 - 12404 chain), member 1

SLC51A Solute carrier family 51, alpha subunit SEQ ID NOS: 12405 - 12409

SLC52A2 Solute carrier family 52 (riboflavin transporter), SEQ ID NOS: 12410 - 12420 member 2

SLC5A6 Solute carrier family 5 (sodium/multivitamin and SEQ ID NOS: 12421 - 12431 iodide cotransporter), member 6

SLC6A9 Solute carrier family 6 (neurotransmitter SEQ ID NOS: 12432 - 12439 transporter, glycine), member 9

SLC8A1 Solute carrier family 8 (sodium/calcium exchanger), SEQ ID NOS: 12440 - 12451 member 1

SLC8B1 Solute carrier family 8 (sodium/lithium/calcium SEQ ID NOS: 12452 - 12462 exchanger), member Bl

SLC9A6 Solute carrier family 9, subfamily A (NHE6, cation SEQ ID NOS: 12463 - 12474 proton antiporter 6), member 6

SLC01A2 Solute carrier organic anion transporter family, SEQ ID NOS: 12475 - 12488 member 1A2

SLIT1 Slit guidance ligand 1 SEQ ID NOS: 12489 - 12492

SLIT2 Slit guidance ligand 2 SEQ ID NOS: 12493 - 12501

SLIT3 Slit guidance ligand 3 SEQ ID NOS: 12502 - 12504

SLITRK3 SLIT and NTRK-like family, member 3 SEQ ID NOS: 12505 - 12507

SLPI Secretory leukocyte peptidase inhibitor SEQ ID NO: 12508

SLTM SAFB-like, transcription modulator SEQ ID NOS: 12509 - 12522

SLURP1 Secreted LY6/PLAUR domain containing 1 SEQ ID NO: 12523

SMARCA2 SWI/SNF related, matrix associated, actin dependent SEQ ID NOS: 12524 - 12571 regulator of chromatin, subfamily a, member 2

SMG6 SMG6 nonsense mediated mRNA decay factor SEQ ID NOS: 12572 - 12583

SMIM7 Small integral membrane protein 7 SEQ ID NOS: 12584 - 12600

SM0C1 SPARC related modular calcium binding 1 SEQ ID NOS: 12601 - 12602

SM0C2 SPARC related modular calcium binding 2 SEQ ID NOS: 12603 - 12607

SMPDL3A Sphingomyelin phosphodiesterase, acid-like 3A SEQ ID NOS: 12608 - 12609

SMPDL3B Sphingomyelin phosphodiesterase, acid-like 3B SEQ ID NOS: 12610 - 12614

SMR3A Submaxillary gland androgen regulated protein 3A SEQ ID NO: 12615

SMR3B Submaxillary gland androgen regulated protein 3B SEQ ID NOS: 12616 - 12618 SNED1 Sushi, nidogen and EGF-like domains 1 SEQID NOS: 12619- 12625

SNTB1 Syntrophin, beta 1 (dystrophin-associated protein SEQID NOS: 12626- 12628

Al, 59kDa, basic component 1)

SNTB2 Syntrophin, beta 2 (dystrophin-associated protein SEQID NOS: 12629- 12633

Al, 59kDa, basic component 2)

SNX14 Sorting nexin 14 SEQID NOS: 12634- 12645

S0D3 Superoxide dismutase 3, extracellular SEQID NOS: 12646- 12647

SOST Sclerostin SEQID NO: 12648

SOSTDC1 Sclerostin domain containing 1 SEQID NOS: 12649- 12650

SOWAHA Sosondowah ankyrin repeat domain family member SEQID NO: 12651

A

SPACA3 Sperm acrosome associated 3 SEQID NOS: 12652 - 12654

SPACA4 Sperm acrosome associated 4 SEQID NO: 12655

SPACA5 Sperm acrosome associated 5 SEQID NOS: 12656- 12657

SPACA5B Sperm acrosome associated 5B SEQID NO: 12658

SPACA7 Sperm acrosome associated 7 SEQID NOS: 12659- 12662

SPAG11A Sperm associated antigen 11A SEQID NOS: 12663- 12671

SPAG11B Sperm associated antigen 11B SEQID NOS: 12672- 12680

SPARC Secreted protein, acidic, cysteine-rich (osteonectin) SEQID NOS: 12681- 12685

SPARCL1 SPARC-like 1 (hevin) SEQID NOS: 12686- 12695

SPATA20 Spermatogenesis associated 20 SEQID NOS: 12696- 12709

SPESP1 Sperm equatorial segment protein 1 SEQID NO: 12710

SPINK1 Serine peptidase inhibitor, Kazal type 1 SEQID NOS: 12711- 12712

SPINK13 Serine peptidase inhibitor, Kazal type 13 (putative) SEQID NOS: 12713 - 12715

SPINK14 Serine peptidase inhibitor, Kazal type 14 (putative) SEQID NOS: 12716- 12717

SPINK2 Serine peptidase inhibitor, Kazal type 2 (acrosin- SEQID NOS: 12718- 12723 trypsin inhibitor)

SPINK4 Serine peptidase inhibitor, Kazal type 4 SEQID NOS: 12724- 12725

SPINK5 Serine peptidase inhibitor, Kazal type 5 SEQID NOS: 12726- 12731

SPINK6 Serine peptidase inhibitor, Kazal type 6 SEQID NOS: 12732 - 12734

SPINK7 Serine peptidase inhibitor, Kazal type 7 (putative) SEQID NOS: 12735 - 12736

SPINK8 Serine peptidase inhibitor, Kazal type 8 (putative) SEQID NO: 12737

SPINK9 Serine peptidase inhibitor, Kazal type 9 SEQID NOS: 12738- 12739

SPINT1 Serine peptidase inhibitor, Kunitz type 1 SEQID NOS: 12740- 12747

SPINT2 Serine peptidase inhibitor, Kunitz type, 2 SEQID NOS: 12748- 12755

SPINT3 Serine peptidase inhibitor, Kunitz type, 3 SEQID NO: 12756

SPINT4 Serine peptidase inhibitor, Kunitz type 4 SEQID NO: 12757

SPOCK1 Sparc/osteonectin, cwcv and kazal-like domains SEQID NOS: 12758- 12761 proteoglycan (testican) 1

SPOCK2 Sparc/osteonectin, cwcv and kazal-like domains SEQID NOS: 12762- 12765 proteoglycan (testican) 2

SPOCK3 Sparc/osteonectin, cwcv and kazal-like domains SEQID NOS: 12766- 12791 proteoglycan (testican) 3

SPON1 Spondin 1, extracellular matrix protein SEQID NO: 12792

SPON2 Spondin 2, extracellular matrix protein SEQID NOS: 12793- 12802

SPP1 Secreted phosphoprotein 1 SEQID NOS: 12803- 12807

SPP2 Secreted phosphoprotein 2, 24kDa SEQID NOS: 12808- 12810 SPRN Shadow of prion protein homolog (zebrafish) SEQID NO: 12811

SPRYD3 SPRY domain containing 3 SEQID NOS: 12812 - 12815

SPRYD4 SPRY domain containing 4 SEQID NO: 12816

SPTY2D1- SPTY2D1 antisense RNA 1 SEQID NOS: 12817- 12822 AS1

SPX Spexin hormone SEQID NOS: 12823 - 12824

SRGN Serglycin SEQID NO: 12825

SRL Sarcalumenin SEQID NOS: 12826- 12828

SRP14 Signal recognition particle 14kDa (homologous Alu SEQID NOS: 12829- 12832

RNA binding protein)

SRPX Sushi-repeat containing protein, X-linked SEQID NOS: 12833 - 12836

SRPX2 Sushi-repeat containing protein, X-linked 2 SEQID NOS: 12837- 12840

SSC4D Scavenger receptor cysteine rich family, 4 domains SEQID NO: 12841

SSC5D Scavenger receptor cysteine rich family, 5 domains SEQID NOS: 12842- 12845

SSPO SCO-spondin SEQID NO: 12846

SSR2 Signal sequence receptor, beta (translocon- SEQID NOS: 12847- 12856 associated protein beta)

SST Somatostatin SEQID NO: 12857

ST3GAL1 ST3 beta-galactoside alpha-2,3-sialyltransferase 1 SEQID NOS: 12858- 12865

ST3GAL4 ST3 beta-galactoside alpha-2,3-sialyltransferase 4 SEQID NOS: 12866- 12881

ST6GAL1 ST6 beta-galactosamide alpha-2,6-sialyltranferase 1 SEQID NOS: 12882- 12897

ST6GALNAC ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl- SEQID NOS: 12898- 12902 2 l,3)-N-acetylgalactosaminide alpha-2,6- sialyltransferase 2

ST6GALNAC ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl- SEQID NOS: 12903 - 12904 5 l,3)-N-acetylgalactosaminide alpha-2,6- sialyltransferase 5

ST6GALNAC ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl- SEQID NOS: 12905- 12912 6 l,3)-N-acetylgalactosaminide alpha-2,6- sialyltransferase 6

ST8SIA2 ST8 alpha-N-acetyl-neuraminide alpha-2,8- SEQID NOS: 12913 - 12915 sialyltransferase 2

ST8SIA4 ST8 alpha-N-acetyl-neuraminide alpha-2,8- SEQID NOS: 12916- 12918 sialyltransferase 4

ST8SIA6 ST8 alpha-N-acetyl-neuraminide alpha-2,8- SEQID NOS: 12919- 12920 sialyltransferase 6

STARD7 StAR-related lipid transfer (START) domain SEQID NOS: 12921 - 12922 containing 7

STATH Statherin SEQID NOS: 12923 - 12925

STC1 Stanniocalcin 1 SEQID NOS: 12926- 12927

STC2 Stanniocalcin 2 SEQID NOS: 12928- 12930

STMND1 Stathmin domain containing 1 SEQID NOS: 12931 - 12932

C7orf73 Chromosome 7 open reading frame 73 SEQID NOS: 12933 - 12934

STOML2 Stomatin (EPB72)-like 2 SEQID NOS: 12935 - 12938

STOX1 Storkhead box 1 SEQID NOS: 12939- 12943

STRC Stereocilin SEQID NOS: 12944-12949

SUCLG1 Succinate-CoA ligase, alpha subunit SEQID NOS: 12950- 12951 SUDS3 SDS3 homolog, SIN3A corepressor complex SEQID NO: 12952 component

SULF1 Sulfatase 1 SEQID NOS: 12953 - 12963

SULF2 Sulfatase 2 SEQID NOS: 12964- 12968

SUMF1 Sulfatase modifying factor 1 SEQID NOS: 12969- 12973

SUMF2 Sulfatase modifying factor 2 SEQID NOS: 12974- 12987

SUSD1 Sushi domain containing 1 SEQID NOS: 12988- 12993

SUSD5 Sushi domain containing 5 SEQID NOS: 12994- 12995

SVEP1 Sushi, von Willebrand factor type A, EGF and SEQID NOS: 12996- 12998 pentraxin domain containing 1

SWSAP1 SWIM-type zinc finger 7 associated protein 1 SEQID NO: 12999

SYAP1 Synapse associated protein 1 SEQID NO: 13000

SYCN Syncollin SEQID NO: 13001

TAC1 Tachykinin, precursor 1 SEQID NOS: 13002 - 13004

TAC3 Tachykinin 3 SEQID NOS: 13005 - 13014

TAC4 Tachykinin 4 (hemokinin) SEQID NOS: 13015 - 13020

TAGLN2 Transgelin 2 SEQID NOS: 13021 - 13024

TAP BP TAP binding protein (tapasin) SEQID NOS: 13025 - 13030

TAP BP L TAP binding protein-like SEQID NOS: 13031 - 13032

TBL2 Transducin (beta)-like 2 SEQID NOS: 13033 - 13045

TBX10 T-box 10 SEQID NO: 13046

TCF12 Transcription factor 12 SEQID NOS: 13047- 13060

TCN1 Transcobalamin 1 (vitamin B12 binding protein, R SEQID NO: 13061

binder family)

TCN2 Transcobalamin II SEQID NOS: 13062 - 13065

TCTN1 Tectonic family member 1 SEQID NOS: 13066- 13084

TCTN3 Tectonic family member 3 SEQID NOS: 13085 - 13089

TDP2 Tyrosyl-DNA phosphodiesterase 2 SEQID NOS: 13090- 13091

C14orf80 Chromosome 14 open reading frame 80 SEQID NOS: 13092 - 13105

TEK TEK tyrosine kinase, endothelial SEQID NOS: 13106- 13110

TEPP Testis, prostate and placenta expressed SEQID NOS: 13111 - 13112

TEX101 Testis expressed 101 SEQID NOS: 13113 - 13114

TEX264 Testis expressed 264 SEQID NOS: 13115 - 13126

Clorf234 Chromosome 1 open reading frame 234 SEQID NOS: 13127- 13129

TF Transferrin SEQID NOS: 13130- 13136

TFAM Transcription factor A, mitochondrial SEQID NOS: 13137- 13139

TFF1 Trefoil factor 1 SEQID NO: 13140

TFF2 Trefoil factor 2 SEQID NO: 13141

TFF3 Trefoil factor 3 (intestinal) SEQID NOS: 13142 - 13144

TFPI Tissue factor pathway inhibitor (lipoprotein- SEQID NOS: 13145 - 13154 associated coagulation inhibitor)

TFPI2 Tissue factor pathway inhibitor 2 SEQID NOS: 13155 - 13156

TG Thyroglobulin SEQID NOS: 13157- 13166

TGFB1 Transforming growth factor, beta 1 SEQID NOS: 13167- 13168

TGFB2 Transforming growth factor, beta 2 SEQID NOS: 13169- 13170

TGFB3 Transforming growth factor, beta 3 SEQID NOS: 13171 - 13172

TGFBI Transforming growth factor, beta-induced, 68kDa SEQID NOS: 13173 - 13180 TGFBR1 Transforming growth factor, beta receptor 1 SEQID NOS: 13181 - 13190

TGFBR3 Transforming growth factor, beta receptor III SEQID NOS: 13191 - 13197

THBS1 Thrombospondin 1 SEQID NOS: 13198- 13199

THBS2 Thrombospondin 2 SEQID NOS: 13200- 13202

THBS3 Thrombospondin 3 SEQID NOS: 13203 - 13207

THBS4 Thrombospondin 4 SEQID NOS: 13208- 13209

TH0C3 THO complex 3 SEQID NOS: 13210- 13219

THPO Thrombopoietin SEQID NOS: 13220- 13225

THSD4 Thrombospondin, type 1, domain containing 4 SEQID NOS: 13226- 13229

THY1 Thy-1 cell surface antigen SEQID NOS: 13230- 13235

TIE1 Tyrosine kinase with immunoglobulin-like and EGF- SEQID NOS: 13236- 13237 like domains 1

TIMMDCl Translocase of inner mitochondrial membrane SEQID NOS: 13238- 13245 domain containing 1

TIMP1 TIMP metallopeptidase inhibitor 1 SEQID NOS: 13246- 13250

TIMP2 TIMP metallopeptidase inhibitor 2 SEQID NOS: 13251 - 13255

TIMP3 TIMP metallopeptidase inhibitor 3 SEQID NO: 13256

TIMP4 TIMP metallopeptidase inhibitor 4 SEQID NO: 13257

TINAGL1 Tubulointerstitial nephritis antigen-like 1 SEQID NOS: 13258- 13260

TINF2 TERF1 (TRFl)-interacting nuclear factor 2 SEQID NOS: 13261 - 13270

TLL2 Tolloid-like 2 SEQID NO: 13271

TLR1 Toll-like receptor 1 SEQID NOS: 13272 - 13277

TLR3 Toll-like receptor 3 SEQID NOS: 13278- 13280

TM2D2 TM2 domain containing 2 SEQID NOS: 13281 - 13286

TM2D3 TM2 domain containing 3 SEQID NOS: 13287- 13294

TM7SF3 Transmembrane 7 superfamily member 3 SEQID NOS: 13295 - 13309

TM9SF1 Transmembrane 9 superfamily member 1 SEQID NOS: 13310- 13320

TMC06 Transmembrane and coiled-coil domains 6 SEQID NOS: 13321 - 13328

TMED1 Transmembrane p24 trafficking protein 1 SEQID NOS: 13329- 13335

TMED2 Transmembrane p24 trafficking protein 2 SEQID NOS: 13336- 13338

TMED3 Transmembrane p24 trafficking protein 3 SEQID NOS: 13339- 13342

TMED4 Transmembrane p24 trafficking protein 4 SEQID NOS: 13343 - 13345

TMED5 Transmembrane p24 trafficking protein 5 SEQID NOS: 13346- 13349

TMED7 Transmembrane p24 trafficking protein 7 SEQID NOS: 13350- 13351

TMED7- TMED7-TICAM2 readthrough SEQID NOS: 13352 - 13353 TICAM2

TMEM108 Transmembrane protein 108 SEQID NOS: 13354- 13362

TMEM116 Transmembrane protein 116 SEQID NOS: 13363 - 13374

TMEM119 Transmembrane protein 119 SEQID NOS: 13375 - 13378

TMEM155 Transmembrane protein 155 SEQID NOS: 13379- 13382

TMEM168 Transmembrane protein 168 SEQID NOS: 13383 - 13388

TMEM178A Transmembrane protein 178A SEQID NOS: 13389- 13390

TMEM179 Transmembrane protein 179 SEQID NOS: 13391 - 13396

TMEM196 Transmembrane protein 196 SEQID NOS: 13397- 13401

TMEM199 Transmembrane protein 199 SEQID NOS: 13402 - 13405

TMEM205 Transmembrane protein 205 SEQID NOS: 13406- 13419

TMEM213 Transmembrane protein 213 SEQID NOS: 13420- 13423 TMEM25 Transmembrane protein 25 SEQID NOS: 13424-13440

TMEM30C Transmembrane protein 30C SEQID NO: 13441

TMEM38B Transmembrane protein 38B SEQID NOS: 13442-13446

TMEM44 Transmembrane protein 44 SEQID NOS: 13447- 13456

TMEM52 Transmembrane protein 52 SEQID NOS: 13457- 13461

TMEM52B Transmembrane protein 52B SEQID NOS: 13462- 13464

TMEM59 Transmembrane protein 59 SEQID NOS: 13465- 13472

TMEM67 Transmembrane protein 67 SEQID NOS: 13473- 13484

TMEM70 Transmembrane protein 70 SEQID NOS: 13485- 13487

TMEM87A Transmembrane protein 87A SEQID NOS: 13488- 13497

TMEM94 Transmembrane protein 94 SEQID NOS: 13498- 13513

TMEM95 Transmembrane protein 95 SEQID NOS: 13514- 13516

TMIGD1 Transmembrane and immunoglobulin domain SEQID NOS: 13517- 13518 containing 1

TMPRSS12 Transmembrane (C-terminal) protease, serine 12 SEQID NOS: 13519- 13520

TMPRSS5 Transmembrane protease, serine 5 SEQID NOS: 13521 - 13532

TMUB1 Transmembrane and ubiquitin-like domain SEQID NOS: 13533 - 13539 containing 1

TMX2 Thioredoxin-related transmembrane protein 2 SEQID NOS: 13540- 13547

TMX3 Thioredoxin-related transmembrane protein 3 SEQID NOS: 13548- 13555

TNC Tenascin C SEQID NOS: 13556- 13564

TNFAIP6 Tumor necrosis factor, alpha-induced protein 6 SEQID NO: 13565

TNFRSF11A Tumor necrosis factor receptor superfamily, SEQID NOS: 13566- 13570 member 11a, NFKB activator

TNFRSF11B Tumor necrosis factor receptor superfamily, SEQID NOS: 13571 - 13572 member lib

TNFRSF12A Tumor necrosis factor receptor superfamily, SEQID NOS: 13573 - 13578 member 12A

TNFRSF14 Tumor necrosis factor receptor superfamily, SEQID NOS: 13579- 13585 member 14

TNFRSF18 Tumor necrosis factor receptor superfamily, SEQID NOS: 13586- 13589 member 18

TNFRSF1A Tumor necrosis factor receptor superfamily, SEQID NOS: 13590- 13598 member 1A

TNFRSF1B Tumor necrosis factor receptor superfamily, SEQID NOS: 13599- 13600 member IB

TNFRSF25 Tumor necrosis factor receptor superfamily, SEQID NOS: 13601 - 13612 member 25

TNFRSF6B Tumor necrosis factor receptor superfamily, SEQID NO: 13613

member 6b, decoy

TNFSF11 Tumor necrosis factor (ligand) superfamily, member SEQID NOS: 13614- 13618

11

TNFSF12 Tumor necrosis factor (ligand) superfamily, member SEQID NOS: 13619- 13620

12

TNFSF12- TNFSF12-TNFSF13 readthrough SEQID NO: 13621 TNFSF13

TNFSF15 Tumor necrosis factor (ligand) superfamily, member SEQID NOS: 13622 - 13623

15

TNN Tenascin N SEQID NOS: 13624- 13626 TNR Tenascin R SEQID NOS: 13627- 13629

TNXB Tenascin XB SEQID NOS: 13630- 13636

FAM179B Family with sequence similarity 179, member B SEQID NOS: 13637- 13642

TOMM7 Translocase of outer mitochondrial membrane 7 SEQID NOS: 13643 - 13646 homolog (yeast)

TOP1MT Topoisomerase (DNA) 1, mitochondrial SEQID NOS: 13647- 13661

TOR1A Torsin family 1, member A (torsin A) SEQID NO: 13662

TO RIB Torsin family 1, member B (torsin B) SEQID NOS: 13663 - 13664

TOR2A Torsin family 2, member A SEQID NOS: 13665 - 13671

TOR3A Torsin family 3, member A SEQID NOS: 13672 - 13676

TPD52 Tumor protein D52 SEQID NOS: 13677- 13689

TPO Thyroid peroxidase SEQID NOS: 13690- 13700

TPP1 Tripeptidyl peptidase 1 SEQID NOS: 13701 - 13718

TPSAB1 Tryptase alpha/beta 1 SEQID NOS: 13719- 13721

TPSB2 Tryptase beta 2 (gene/pseudogene) SEQID NOS: 13722 - 13724

TPSD1 Tryptase delta 1 SEQID NOS: 13725 - 13726

TPST1 Tyrosylprotein sulfotransferase 1 SEQID NOS: 13727- 13729

TPST2 Tyrosylprotein sulfotransferase 2 SEQID NOS: 13730- 13738

TRABD2A TraB domain containing 2A SEQID NOS: 13739- 13741

TRABD2B TraB domain containing 2B SEQID NO: 13742

TREH Trehalase (brush-border membrane glycoprotein) SEQID NOS: 13743 - 13745

TREM1 Triggering receptor expressed on myeloid cells 1 SEQID NOS: 13746- 13749

TREM2 Triggering receptor expressed on myeloid cells 2 SEQID NOS: 13750- 13752

TRH Thyrotropin-releasing hormone SEQID NOS: 13753 - 13754

TRIM24 Tripartite motif containing 24 SEQID NOS: 13755 - 13756

TRIM28 Tripartite motif containing 28 SEQID NOS: 13757- 13762

TRIO Trio Rho guanine nucleotide exchange factor SEQID NOS: 13763 - 13769

TRNP1 TMFl-regulated nuclear protein 1 SEQID NOS: 13770- 13771

TSC22D4 TSC22 domain family, member 4 SEQID NOS: 13772 - 13775

TSHB Thyroid stimulating hormone, beta SEQID NOS: 13776- 13777

TSHR Thyroid stimulating hormone receptor SEQID NOS: 13778- 13785

TSKU Tsukushi, small leucine rich proteoglycan SEQID NOS: 13786- 13790

TSLP Thymic stromal lymphopoietin SEQID NOS: 13791 - 13793

TSPAN3 Tetraspanin 3 SEQID NOS: 13794- 13799

TSPAN31 Tetraspanin 31 SEQID NOS: 13800- 13806

TSPEAR Thrombospondin-type laminin G domain and EAR SEQID NOS: 13807- 13810 repeats

TTC13 Tetratricopeptide repeat domain 13 SEQID NOS: 13811 - 13817

TTC19 Tetratricopeptide repeat domain 19 SEQID NOS: 13818- 13823

TTC9B Tetratricopeptide repeat domain 9B SEQID NO: 13824

TTLLll Tubulin tyrosine ligase-like family member 11 SEQID NOS: 13825 - 13829

TTR Transthyretin SEQID NOS: 13830- 13832

TWSG1 Twisted gastrulation BMP signaling modulator 1 SEQID NOS: 13833- 13835

TXNDC12 Thioredoxin domain containing 12 (endoplasmic SEQID NOS: 13836- 13838 reticulum)

TXNDC15 Thioredoxin domain containing 15 SEQID NOS: 13839- 13845 TXNDC5 Thioredoxin domain containing 5 (endoplasmic SEQID NOS: 13846- 13847 reticulum)

TXNRD2 Thioredoxin reductase 2 SEQID NOS: 13848- 13860

TYRP1 Tyrosinase-related protein 1 SEQID NOS: 13861- 13863

UBAC2 UBA domain containing 2 SEQID NOS: 13864- 13868

UBALD1 UBA-like domain containing 1 SEQID NOS: 13869- 13877

UBAP2 Ubiquitin associated protein 2 SEQID NOS: 13878- 13884

UBXN8 UBX domain protein 8 SEQID NOS: 13885- 13891

UCMA Upper zone of growth plate and cartilage matrix SEQID NOS: 13892 - 13893 associated

UCN Urocortin SEQID NO: 13894

UCN2 Urocortin 2 SEQID NO: 13895

UCN3 Urocortin 3 SEQID NO: 13896

UGGT2 UDP-glucose glycoprotein glucosyltransferase 2 SEQID NOS: 13897- 13902

UGT1A10 UDP glucuronosyltransferase 1 family, polypeptide SEQID NOS: 13903- 13904

A10

UGT2A1 UDP glucuronosyltransferase 2 family, polypeptide SEQID NOS: 13905- 13909

Al, complex locus

UGT2B11 UDP glucuronosyltransferase 2 family, polypeptide SEQID NO: 13910

Bll

UGT2B28 UDP glucuronosyltransferase 2 family, polypeptide SEQID NOS: 13911- 13912

B28

UGT2B4 UDP glucuronosyltransferase 2 family, polypeptide SEQID NOS: 13913 - 13916

B4

UGT2B7 UDP glucuronosyltransferase 2 family, polypeptide SEQID NOS: 13917- 13920

B7

UGT3A1 UDP glycosyltransferase 3 family, polypeptide Al SEQID NOS: 13921 - 13926

UGT3A2 UDP glycosyltransferase 3 family, polypeptide A2 SEQID NOS: 13927- 13930

UGT8 UDP glycosyltransferase 8 SEQID NOS: 13931 - 13933

ULBP3 UL16 binding protein 3 SEQID NOS: 13934- 13935

UMOD Uromodulin SEQID NOS: 13936- 13947

UNC5C Unc-5 netrin receptor C SEQID NOS: 13948- 13952

UPK3B Uroplakin 3B SEQID NOS: 13953 - 13955

USP11 Ubiquitin specific peptidase 11 SEQID NOS: 13956- 13959

USP14 Ubiquitin specific peptidase 14 (tRNA-guanine SEQID NOS: 13960- 13966 transglycosylase)

USP3 Ubiquitin specific peptidase 3 SEQID NOS: 13967- 13982

CIRH1A Cirrhosis, autosomal recessive 1A (cirhin) SEQID NOS: 13983 - 13992

UTS2 Urotensin 2 SEQID NOS: 13993- 13995

UTS2B Urotensin 2B SEQID NOS: 13996- 14001

UTY Ubiquitously transcribed tetratricopeptide repeat SEQID NOS: 14002 - 14014 containing, Y-linked

UXS1 UDP-glucuronate decarboxylase 1 SEQID NOS: 14015- 14022

VASH1 Vasohibin 1 SEQID NOS: 14023- 14025

VCAN Versican SEQID NOS: 14026- 14032

VEGFA Vascular endothelial growth factor A SEQID NOS: 14033- 14058

VEGFB Vascular endothelial growth factor B SEQID NOS: 14059- 14061

VEGFC Vascular endothelial growth factor C SEQID NO: 14062 FIGF C-fos induced growth factor (vascular endothelial SEQID NO: 14063 growth factor D)

VGF VGF nerve growth factor inducible SEQID NOS: 14064-14066

VIP Vasoactive intestinal peptide SEQID NOS: 14067-14069

VIPR2 Vasoactive intestinal peptide receptor 2 SEQID NOS: 14070-14073

VIT Vitrin SEQID NOS: 14074-14081

VK0RC1 Vitamin K epoxide reductase complex, subunit 1 SEQID NOS: 14082- 14089

VLDLR Very low density lipoprotein receptor SEQID NOS: 14090-14092

VMOl Vitelline membrane outer layer 1 homolog (chicken) SEQID NOS: 14093- 14096

VNN1 Vanin 1 SEQID NO: 14097

VNN2 Vanin 2 SEQID NOS: 14098-14111

VNN3 Vanin 3 SEQID NOS: 14112-14123

VOPP1 Vesicular, overexpressed in cancer, prosurvival SEQID NOS: 14124-14136 protein 1

VPREB1 Pre-B lymphocyte 1 SEQID NOS: 14137-14138

VPREB3 Pre-B lymphocyte 3 SEQID NOS: 14139-14140

VPS37B Vacuolar protein sorting 37 homolog B (S. cerevisiae) SEQID NOS: 14141-14143

VPS51 Vacuolar protein sorting 51 homolog (S. cerevisiae) SEQID NOS: 14144-14155

VSIG1 V-set and immunoglobulin domain containing 1 SEQID NOS: 14156-14158

VSIG10 V-set and immunoglobulin domain containing 10 SEQID NOS: 14159-14160

VSTM1 V-set and transmembrane domain containing 1 SEQID NOS: 14161-14167

VSTM2A V-set and transmembrane domain containing 2A SEQID NOS: 14168-14171

VSTM2B V-set and transmembrane domain containing 2B SEQID NO: 14172

VSTM2L V-set and transmembrane domain containing 2 like SEQID NOS: 14173-14175

VSTM4 V-set and transmembrane domain containing 4 SEQID NOS: 14176-14177

VTN Vitronectin SEQID NOS: 14178-14179

VWA1 Von Willebrand factor A domain containing 1 SEQID NOS: 14180-14183

VWA2 Von Willebrand factor A domain containing 2 SEQID NOS: 14184-14185

VWA5B2 Von Willebrand factor A domain containing 5B2 SEQID NOS: 14186-14187

VWA7 Von Willebrand factor A domain containing 7 SEQID NO: 14188

VWC2 Von Willebrand factor C domain containing 2 SEQID NO: 14189

VWC2L Von Willebrand factor C domain containing protein SEQID NOS: 14190-14191

2-like

VWCE Von Willebrand factor C and EGF domains SEQID NOS: 14192-14196

VWDE Von Willebrand factor D and EGF domains SEQID NOS: 14197-14202

VWF Von Willebrand factor SEQID NOS: 14203- 14205

WDR25 WD repeat domain 25 SEQID NOS: 14206- 14212

WDR81 WD repeat domain 81 SEQID NOS: 14213- 14222

WDR90 WD repeat domain 90 SEQID NOS: 14223- 14230

WFDC1 WAP four-disulfide core domain 1 SEQID NOS: 14231- 14233

WFDCIOA WAP four-disulfide core domain 10A SEQID NO: 14234

WFDCIOB WAP four-disulfide core domain 10B SEQID NOS: 14235- 14236

WFDC11 WAP four-disulfide core domain 11 SEQID NOS: 14237- 14239

WFDC12 WAP four-disulfide core domain 12 SEQID NO: 14240

WFDC13 WAP four-disulfide core domain 13 SEQID NO: 14241

WFDC2 WAP four-disulfide core domain 2 SEQID NOS: 14242-14246

WFDC3 WAP four-disulfide core domain 3 SEQID NOS: 14247- 14250 WFDC5 WAP four-disulfide core domain 5 SEQIDNOS: 14251- 14252

WFDC6 WAP four-disulfide core domain 6 SEQIDNOS: 14253- 14254

WFDC8 WAP four-disulfide core domain 8 SEQIDNOS: 14255- 14256

WFIKKN1 WAP, follistatin/kazal, immunoglobulin, kunitz and SEQID NO: 14257

netrin domain containing 1

WFIKKN2 WAP, follistatin/kazal, immunoglobulin, kunitz and SEQID NOS: 14258- 14259 netrin domain containing 2

DFNB31 Deafness, autosomal recessive 31 SEQID NOS: 14260- 14263

WIF1 WNT inhibitory factor 1 SEQID NOS: 14264-14266

WISP1 WNT1 inducible signaling pathway protein 1 SEQID NOS: 14267- 14271

WISP2 WNT1 inducible signaling pathway protein 2 SEQIDNOS: 14272- 14274

WISP3 WNT1 inducible signaling pathway protein 3 SEQIDNOS: 14275- 14282

WNK1 WNK lysine deficient protein kinase 1 SEQIDNOS: 14283- 14296

WNT1 Wingless-type MMTV integration site family, SEQID NOS: 14297- 14298 member 1

WNT10B Wingless-type MMTV integration site family, SEQID NOS: 14299- 14303 member 10B

WNT11 Wingless-type MMTV integration site family, SEQID NOS: 14304-14306 member 11

WNT16 Wingless-type MMTV integration site family, SEQID NOS: 14307- 14308 member 16

WNT2 Wingless-type MMTV integration site family SEQID NOS: 14309- 14311 member 2

WNT3 Wingless-type MMTV integration site family, SEQID NO: 14312

member 3

WNT3A Wingless-type MMTV integration site family, SEQID NO: 14313

member 3A

WNT5A Wingless-type MMTV integration site family, SEQID NOS: 14314-14317 member 5A

WNT5B Wingless-type MMTV integration site family, SEQID NOS: 14318- 14324 member 5B

WNT6 Wingless-type MMTV integration site family, SEQID NO: 14325

member 6

WNT7A Wingless-type MMTV integration site family, SEQID NO: 14326

member 7A

WNT7B Wingless-type MMTV integration site family, SEQID NOS: 14327- 14331 member 7B

WNT8A Wingless-type MMTV integration site family, SEQID NOS: 14332 - 14335 member 8A

WNT8B Wingless-type MMTV integration site family, SEQID NO: 14336

member 8B

WNT9A Wingless-type MMTV integration site family, SEQID NO: 14337

member 9A

WNT9B Wingless-type MMTV integration site family, SEQID NOS: 14338- 14340 member 9B

WSB1 WD repeat and SOCS box containing 1 SEQIDNOS: 14341-14350

WSCD1 WSC domain containing 1 SEQIDNOS: 14351- 14360

WSCD2 WSC domain containing 2 SEQIDNOS: 14361- 14364

XCL1 Chemokine (C motif) ligand 1 SEQID NO: 14365 XCL2 Chemokine (C motif) ligand 2 SEQID NO: 14366

XPNPEP2 X-prolyl aminopeptidase (aminopeptidase P) 2, SEQID NOS: 14367- 14368 membrane-bound

XXYLT1 Xyloside xylosyltransferase 1 SEQID NOS: 14369- 14374

XYLT1 Xylosyltransferase 1 SEQID NO: 14375

XYLT2 Xylosyltransferase II SEQID NOS: 14376- 14381

ZFYVE21 Zinc finger, FYVE domain containing 21 SEQID NOS: 14382- 14386

ZG16 Zymogen granule protein 16 SEQID NO: 14387

ZG16B Zymogen granule protein 16B SEQID NOS: 14388- 14391

ZIC4 Zic family member 4 SEQID NOS: 14392-14400

ZNF207 Zinc finger protein 207 SEQID NOS: 14401-14411

ZNF26 Zinc finger protein 26 SEQID NOS: 14412-14415

ZNF34 Zinc finger protein 34 SEQID NOS: 14416-14419

ZNF419 Zinc finger protein 419 SEQID NOS: 14420-14434

ZNF433 Zinc finger protein 433 SEQID NOS: 14435-14444

ZNF449 Zinc finger protein 449 SEQID NOS: 14445-14446

ZNF488 Zinc finger protein 488 SEQID NOS: 14447-14448

ZNF511 Zinc finger protein 511 SEQID NOS: 14449-14450

ZNF570 Zinc finger protein 570 SEQID NOS: 14451-14456

ZNF691 Zinc finger protein 691 SEQID NOS: 14457-14464

ZNF98 Zinc finger protein 98 SEQID NOS: 14465-14468

ZPBP Zona pellucida binding protein SEQID NOS: 14469-14472

ZPBP2 Zona pellucida binding protein 2 SEQID NOS: 14473-14476

ZSCAN29 Zinc finger and SCAN domain containing 29 SEQID NOS: 14477-14483

[0399] In certain embodiments, the therapeutic protein is not secreted, but rather functions intracellularly.

[0400] In certain embodiments, the therapeutic protein is not secreted, but rather directs a modified cell of the disclosure to a cell niche of a subject's body.

[0401] In certain embodiments of the methods of the disclosure, the subject has a disease or disorder and the plurality of therapeutic immune cells or immune precursor cells improves a sign or symptom of the disease or disorder, optionally by providing a therapeutic protein systemically or locally within the subject that acts upon the immune cell, the immune precursor cell or a second cell in the subject. Exemplary therapeutic secreted proteins may be used as a monotherapy or in combination with another therapy in the treatment or prevention of any disease or disorder. These secreted proteins may be used as a monotherapy or in combination with another therapy for enzyme replacement and/or administration of biologic therapeutics.

Inducible Proapoptotic Polypeptides [0402] Inducible proapoptotic polypeptides of the disclosure are superior to existing inducible polypeptides because the inducible proapoptotic polypeptides of the disclosure are far less immunogenic. While inducible proapoptotic polypeptides of the disclosure are recombinant polypeptides, and, therefore, non-naturally occurring, the sequences that are recombined to produce the inducible proapoptotic polypeptides of the disclosure do not comprise non-human sequences that the host human immune system could recognize as "non-self and, consequently, induce an immune response in the subject receiving an inducible proapoptotic polypeptide of the disclosure, a cell comprising the inducible proapoptotic polypeptide or a composition comprising the inducible proapoptotic polypeptide or the cell comprising the inducible proapoptotic polypeptide.

[0403] Modified cells and/or transposons of the disclosure may comprise an inducible proapoptotic polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a proapoptotic polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments, the non-human sequence comprises a restriction site. In certain embodiments, the ligand binding region may be a multimeric ligand binding region. Inducible proapoptotic polypeptides of the disclosure may also be referred to as an "iC9 safety switch". In certain embodiments, modified cells and/or transposons of the disclosure may comprise an inducible caspase polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a caspase polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments, modified cells and/or transposons of the disclosure may comprise an inducible caspase polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a caspase polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments, transposons of the disclosure may comprise an inducible caspase polypeptide comprising (a) a ligand binding region, (b) a linker, and (c) a truncated caspase 9 polypeptide, wherein the inducible proapoptotic polypeptide does not comprise a non-human sequence. In certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the ligand binding region may comprise a FK506 binding protein 12 (FKBP12) polypeptide. In certain embodiments, the amino acid sequence of the ligand binding region that comprise a FK506 binding protein 12 (FKBP 12) polypeptide may comprise a modification at position 36 of the sequence. The modification may be a substitution of valine (V) for phenylalanine (F) at position 36 (F36V).

[0404] In certain embodiments, the FKBP 12 polypeptide is encoded by an amino acid sequence comprising GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKPFKFMLGKQEVI RGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLKLE (SEQ ID NO: 14635).

[0405] In certain embodiments, the FKBP12 polypeptide is encoded by a nucleic acid sequence comprising

GGGGTCCAGGTCGAGACTATTTCACCAGGGGATGGGCGAACATTTCCAAAAAGG GGCCAGACTTGCGTCGTGCATTACACCGGGATGCTGGAGGACGGGAAGAAAGTG GACAGCTCCAGGGATCGCAACAAGCCCTTCAAGTTCATGCTGGGAAAGCAGGAA GTGATCCGAGGATGGGAGGAAGGCGTGGCACAGATGTCAGTCGGCCAGCGGGCC AAACTGACCATTAGCCCTGACTACGCTTATGGAGCAACAGGCCACCCAGGGATC ATTCCCCCTCATGCCACCCTGGTCTTCGAT GTGGAACTGCTGAAGCTGGAG (SEQ ID NO: 14636). In certain embodiments, the induction agent specific for the ligand binding region may comprise a FK506 binding protein 12 (FKBP12) polypeptide having a substitution of valine (V) for phenylalanine (F) at position 36 (F36V) comprises AP20187 and/or AP1903, both synthetic drugs.

[0406] In certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the linker region is encoded by an amino acid comprising GGGGS (SEQ ID NO: 14637) or a nucleic acid sequence comprising GGAGGAGGAGGATCC (SEQ ID NO: 14638). In certain

embodiments, the nucleic acid sequence encoding the linker does not comprise a restriction site.

[0407] In certain embodiments of the truncated caspase 9 polypeptides of the disclosure, the truncated caspase 9 polypeptide is encoded by an amino acid sequence that does not comprise an arginine (R) at position 87 of the sequence. Alternatively, or in addition, in certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the truncated caspase 9 polypeptide is encoded by an amino acid sequence that does not comprise an alanine (A) at position 282 the sequence. In certain embodiments of the inducible proapoptotic polypeptides, inducible caspase polypeptides or truncated caspase 9 polypeptides of the disclosure, the truncated caspase 9 polypeptide is encoded by an amino acid comprising

GFGDVGALESLRGNADLAYILSMEPCGHCLIINNVNFCRESGLRTRTGSNIDCEKLRR RFSSLHFMVEVKGDLTAKKMVLALLELAQQDHGALDCCVVVILSHGCQASHLQFPG AVYGTDGCPVSVEKIVNIFNGTSCPSLGGKPKLFFIQACGGEQKDHGFEVASTSPEDE SPGSNPEPDATPFQEGLRTFDQLDAISSLPTPSDIFVSYSTFPGFVSWRDPKSGSWYVE TLDDIFEQWAHSEDLQSLLLRVANAVSVKGIYKQMPGCFNFLRKKLFFKTS (SEQ ID NO: 14639) or a nucleic acid sequence comprising

TTTGGGGACGTGGGGGCCCTGGAGTCTCTGCGAGGAAATGCCGATCTGGCTTACA

TCCTGAGCATGGAACCCTGCGGCCACTGTCTGATCATTAACAATGTGAACTTCTG

CAGAGAAAGCGGACTGCGAACACGGACTGGCTCCAATATTGACTGTGAGAAGCT

GCGGAGAAGGTTCTCTAGTCTGCACTTTATGGTCGAAGTGAAAGGGGATCTGACC

GCCAAGAAAATGGTGCTGGCCCTGCTGGAGCTGGCTCAGCAGGACCATGGAGCT

CTGGATTGCTGCGTGGTCGTGATCCTGTCCCACGGGTGCCAGGCTTCTCATCTGC

AGTTCCCCGGAGCAGTGTACGGAACAGACGGCTGTCCTGTCAGCGTGGAGAAGA

TCGTCAACATCTTCAACGGCACTTCTTGCCCTAGTCTGGGGGGAAAGCCAAAACT

GTTCTTTATCCAGGCCTGTGGCGGGGAACAGAAAGATCACGGCTTCGAGGTGGC

CAGCACCAGCCCTGAGGACGAATCACCAGGGAGCAACCCTGAACCAGATGCAAC

TCCATTCCAGGAGGGACTGAGGACCTTTGACCAGCTGGATGCTATCTCAAGCCTG

CCCACTCCTAGTGACATTTTCGTGTCTTACAGTACCTTCCCAGGCTTTGTCTCATG

GCGCGATCCCAAGTCAGGGAGCTGGTACGTGGAGACACTGGACGACATCTTTGA

ACAGTGGGCCCATTCAGAGGACCTGCAGAGCCTGCTGCTGCGAGTGGCAAACGC

TGTCTCTGTGAAGGGCATCTACAAACAGATGCCCGGGTGCTTCAATTTTCTGAGA

AAGAAACTGTTCTTTAAGACTTCC (SEQ ID NO: 14640).

[0408] In certain embodiments of the inducible proapoptotic polypeptides, wherein the polypeptide comprises a truncated caspase 9 polypeptide, the inducible proapoptotic polypeptide is encoded by an amino acid sequence comprising

GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKPFKFMLGKQEVI RGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELLKLEGGGGSG FGDVGALESLRGNADLAYILSMEPCGHCLIINNVNFCRESGLRTRTGSNIDCEKLRRR FSSLHFMVEVKGDLTAKKMVLALLELAQQDHGALDCCVVVILSHGCQASHLQFPGA VYGTDGCPVSVEKIVNIFNGTSCPSLGGKPKLFFIQACGGEQKDHGFEVASTSPEDES PGSNPEPDATPFQEGLRTFDQLDAISSLPTPSDIFVSYSTFPGFVSWRDPKSGSWYVET LDDIFEQWAHSEDLQSLLLRVANAVSVKGIYKQMPGCFNFLRKKLFFKTS (SEQ ID NO: 14641) or the nucleic acid sequence comprising

ggggtccaggtcgagactatttcaccaggggatgggcgaacatttccaaaaaggggc cagacttgcgtcgtgcattacaccgggatg ctggaggacgggaagaaagtggacagctccagggatcgcaacaagcccttcaagttcatg ctgggaaagcaggaagtgatccgag gatgggaggaaggcgtggcacagatgtcagtcggccagcgggccaaactgaccattagcc ctgactacgcttatggagcaacagg ccacccagggatcattccccctcatgccaccctggtcttcgatgtggaactgctgaagct ggagggaggaggaggatccggatttgg ggacgtgggggccctggagtctctgcgaggaaatgccgatctggcttacatcctgagcat ggaaccctgcggccactgtctgatcatt aacaatgtgaacttctgcagagaaagcggactgcgaacacggactggctccaatattgac tgtgagaagctgcggagaaggttctcta gtctgcactttatggtcgaagtgaaaggggatctgaccgccaagaaaatggtgctggccc tgctggagctggctcagcaggaccatg gagctctggattgctgcgtggtcgtgatcctgtcccacgggtgccaggcttctcatctgc agttccccggagcagtgtacggaacagac ggctgtcctgtcagcgtggagaagatcgtcaacatcttcaacggcacttcttgccctagt ctggggggaaagccaaaactgttctttatc caggcctgtggcggggaacagaaagatcacggcttcgaggtggccagcaccagccctgag gacgaatcaccagggagcaaccct gaaccagatgcaactccattccaggagggactgaggacc†atgaccagctggatgcta tctcaagcctgcccactcctagtgaca†ottc gtgtcttacagtaccttcccaggCTOtgtctcatggcgcgatcccaagtcagggagctgg tacgtggagacactggacgacatctttgaa cagtgggcccattcagaggacctgcagagcctgctgctgcgagtggcaaacgctgtctct gtgaagggcatctacaaacagatgccc gggtgcttcaattttctgagaaagaaactgttctttaagacttcc (SEQ ID NO: 14642).

Construct Elements

[0409] Transposons and other delivery vectors of the disclosure may comprise at least one self-cleaving peptide(s) located, for example, between one or more of a sequence encoding an inducible proapoptotic polypeptide of the disclosure, a sequence encoding a therapeutic protein of the disclosure and a selection gene of the disclosure.

[0410] Transposons and other delivery vectorsof the disclosure may comprise at least two self-cleaving peptide(s), a first self-cleaving peptide located, for example, upstream or immediately upstream of an inducible proapoptotic polypeptide of the disclosure of the disclosure and a second first self-cleaving peptide located, for example, downstream or immediately upstream of an inducible proapoptotic polypeptide of the disclosure of the disclosure.

[0411] The at least one self-cleaving peptide may comprise, for example, a T2A peptide, GSG-T2A peptide, an E2A peptide, a GSG-E2A peptide, an F2A peptide, a GSG-F2A peptide, a P2A peptide, or a GSG-P2A peptide. A T2A peptide may comprise an amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643). A GSG-T2A peptide may comprise an amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644). A GSG-T2A peptide may comprise a nucleic acid sequence comprising

ggatctggagagggaaggggaagcctgctgacctgtggagacgtggaggaaaaccca ggacca (SEQ ID NO: 14645). An E2A peptide may comprise an amino acid sequence comprising

QCTNYALLKLAGDVESNPGP (SEQ ID NO: 14646) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising QCTNYALLKLAGDVESNPGP (SEQ ID NO: 14646). A GSG-E2A peptide may comprise an amino acid sequence comprising GSGQCTNYALLKLAGDVESNPGP (SEQ ID NO: 14647) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGQCTNYALLKLAGDVESNPGP (SEQ ID NO: 14647). An F2A peptide may comprise an amino acid sequence comprising

VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14648) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14648). A GSG-F2A peptide may comprise an amino acid sequence comprising GSGVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14649) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14649). A P2A peptide may comprise an amino acid sequence comprising

ATNFSLLKQAGDVEENPGP (SEQ ID NO: 14650) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

ATNFSLLKQAGDVEENPGP (SEQ ID NO: 14650). A GSG-P2A peptide may comprise an amino acid sequence comprising GS GATNF SLLKQ AGD VEENP GP (SEQ ID NO: 14651) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 14651).

[0412] Transposons and other delivery vectors of the disclosure may comprise a first and a second self-cleaving peptide, the first self-cleaving peptide located, for example, upstream of one or more of a sequence encoding a therapeutic protein of the disclosure the second self- cleaving peptide located, for example, downstream of a sequence encoding a therapeutic protein of the disclosure. The first and/or the second self-cleaving peptide may comprise, for example, a T2A peptide, GSG-T2A peptide, an E2A peptide, a GSG-E2A peptide, an F2A peptide, a GSG-F2A peptide, a P2A peptide, or a GSG-P2A peptide. A T2A peptide may comprise an amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising EGRGSLLTCGDVEENPGP (SEQ ID NO: 14643). A GSG-T2A peptide may comprise an amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGEGRGSLLTCGDVEENPGP (SEQ ID NO: 14644). A GSG-T2A peptide may comprise a nucleic acid sequence comprising

ggatctggagagggaaggggaagcctgctgacctgtggagacgtggaggaaaaccca ggacca (SEQ ID NO: 14645). An E2A peptide may comprise an amino acid sequence comprising QCTNYALLKLAGDVESNPGP (SEQ ID NO: 14646) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

QCTNYALLKLAGDVESNPGP (SEQ ID NO: 14646). A GSG-E2A peptide may comprise an amino acid sequence comprising GSGQCTNYALLKLAGDVESNPGP (SEQ ID NO: 14647) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGQCTNYALLKLAGDVESNPGP (SEQ ID NO: 14647). An F2A peptide may comprise an amino acid sequence comprising

VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14648) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

VKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14648) . A GSG-F2A peptide may comprise an amino acid sequence comprising GSGVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14649) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGVKQTLNFDLLKLAGDVESNPGP (SEQ ID NO: 14649). A P2A peptide may comprise an amino acid sequence comprising

ATNFSLLKQAGDVEENPGP (SEQ ID NO: 14650) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising

ATNFSLLKQAGDVEENPGP (SEQ ID NO: 14650). A GSG-P2A peptide may comprise an amino acid sequence comprising GS GATNF SLLKQ AGD VEENP GP (SEQ ID NO: 14651) or a sequence having at least 70%, 80%, 90%, 95%, or 99% identity to the amino acid sequence comprising GSGATNFSLLKQAGDVEENPGP (SEQ ID NO: 14651).

[0413] Transposons of the disclosure may comprise a selection gene. The selection gene may encode a gene product essential for cell viability and survival. The selection gene may encode a gene product essential for cell viability and survival when challenged by selective cell culture conditions. Selective cell culture conditions may comprise a compound harmful to cell viability or survival and wherein the gene product confers resistance to the compound.

[0414] By "stable transformation" is intended that the polynucleotide construct introduced into a cell integrates into the genome of the host and is capable of being inherited by progeny thereof.

[0415] By "transient transformation" is intended that a polynucleotide construct introduced into the host does not integrate into the genome of the host.

[0416] All percentages and ratios are calculated based on the total composition unless otherwise indicated.

[0417] Every maximum numerical limitation given throughout this disclosure includes every lower numerical limitation, as if such lower numerical limitations were expressly written herein. Every minimum numerical limitation given throughout this disclosure will include every higher numerical limitation, as if such higher numerical limitations were expressly written herein. Every numerical range given throughout this disclosure will include every narrower numerical range that falls within such broader numerical range, as if such narrower numerical ranges were all expressly written herein.

[0418] The values disclosed herein are not to be understood as being strictly limited to the exact numerical values recited. Instead, unless otherwise specified, each such value is intended to mean both the recited value and a functionally equivalent range surrounding that value. For example, a value disclosed as "20 μιτι" is intended to mean "about 20 μιτι."

[0419] Every document cited herein, including any cross referenced or related patent or application, is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern.

[0420] While particular embodiments of the disclosure have been illustrated and described, various other changes and modifications can be made without departing from the spirit and scope of the disclosure. The scope of the appended claims includes all such changes and modifications that are within the scope of this disclosure.

EXAMPLES

[0421] In order that the invention disclosed herein may be more efficiently understood, examples are provided below. It should be understood that these examples are for illustrative purposes only and are not to be construed as limiting the invention in any manner.

Throughout these examples, molecular cloning reactions, and other standard recombinant DNA techniques, were carried out according to methods described in Maniatis et al, Molecular Cloning - A Laboratory Manual, 2nd ed., Cold Spring Harbor Press (1989), using commercially available reagents, except where otherwise noted.

EXAMPLE 1: Ex vivo genetic modification of T cells [0422] The piggyBac™ (PB) transposon system was used for genetically modifying human lymphocytes for production of autologous CAR-T immunotherapies and other applications. T Lymphocytes purified from patient blood or apheresis product was electroporated with a plasmid DNA transposon and a transposase. Several different electroporation systems have been used for T cell delivery of the transposon system, including the Neon (Thermo Fisher), BTX ECM 830 (Harvard Apparatus), Gene Pulser (BioRad), MaxCyte PulseAgile

(MaxCyte), and the Amaxa 2B and Amaxa 4D (Lonza). Some were tested using

manufacturer provided or recommended electroporation buffer, as well as several in-house developed buffers. Results were consistent with the prevailing dogma that resting T lymphocytes are particularly refractory to DNA transfection and that there appeared to be an inverse relationship between electroporation efficiency, as measured by GFP expression from the electroporated plasmid, and cell viability. Figure 1 shows an example of an experiment testing multiple electroporation systems and nucleofection programs.

[0423] To further test whether or not plasmid DNA was toxic to T cells during nucleofection, primary human T lymphocytes were electroporated with two different DNA plasmids. The first plasmid was a pmaxGFP™ plasmid that is provided as a control plasmid in the Lonza Amaxa nucleofection kit. It is highly purified by HPLC and does not contain endotoxin at detectable levels. The second plasmid was our in-house produced PB transposon encoding a human EF1 alpha promoter driving GFP. Transfection efficiency, as measured by GFP expression from the electroporated plasmid, and cell viability was assessed by FACS at days 2, 3, and 6 post-electroporation. Data are displayed in Figure 2. While mock electroporated cells (no plasmid DNA) exhibited relatively high levels of cell viability by day 6 post- electroporation, 54%, T cells electroporated with either plasmid were only 1.4-2.6% viable. These data show that plasmid DNA was cytotoxic to T lymphocytes. In addition, these data show that DNA-mediated toxicity was not due to transposon element such as the ITR regions or the core insulators since the pmaxGFP™ plasmid are devoid of these elements and was also cytotoxic at the same DNA concentration. Both plasmids are approximately the same size, meaning that similar amounts of DNA were electroporated into the T cells.

[0424] To test whether or not DNA-mediated toxicity in T cells was dose dependent, we performed a titration of our PB-GFP plasmid. Figure 3 shows that as the dose of plasmid DNA added to the nucleofection reaction was increased incrementally (1.3, 2.5, 5.0, 10.0, and 20.0 μg of plasmid DNA), cell viability decreased as measured at both day 1 and 5 post- nucleofection. Even 1.3 μg of plasmid DNA was responsible for a 2.4-fold decrease in T cell viability by day 4. [0425] Since it was clear that plasmid DNA is toxic to T cells during nucleofection, we considered whether or not extracellular plasmid DNA was contributing to cell death. Figure 4 shows that extracellular plasmid DNA was not cytotoxic to T cells. In that experiment, 5 μg of plasmid DNA was added to the cells 45 min post-electroporation and little cell death was observed at day 1 or day 4. Similarly, when 5 μg of plasmid DNA was added to the nucleofection reaction in the absence of electroporation, little cell death was observed.

However, when the plasmid DNA was added before the electroporation reaction, the cells exhibited a 2.0-fold reduction in cell viability at day 1 and a 13.2-fold reduction at day 4.

[0426] Since DNA-mediated toxicity is dose dependent, we next focused our attention on ways to reduce the total amount of DNA delivered to the T cells that is required for transposition. One relatively straightforward way of achieving this would be to deliver the transposase as encoded in mRNA instead of encoded in DNA. mRNA delivery to primary human T cells is very efficient, resulting in high transfection efficiency and high viability. We subcloned the Super piggyBac™ (SPB) transposase enzyme into our in-house mRNA production vector and produced high quality SPB mRNA. Co-delivery of PB-GFP transposon with various doses of SPB mRNA (30, 10, 3.3, 3, 1, 0.33 μg mRNA) in Jurkat cells demonstrated strong transposition at all doses tested (Figure 5). These data show that SPB transposase can be delivered and are equally effective as either plasmid DNA or mRNA. In addition, that the amount of SPB mRNA makes little difference in overall transposition efficiency in Jurkats, in either overall percentage of GFP+ cells or in the MFI of GFP expression. To see if this also holds true for T lymphocytes, we delivered PB-GFP with either SPB plasmid DNA, at a 3: 1 ratio, or 5 μg of SPB mRNA. Seven (7) days following the nucleofection reaction and the addition of IL7 and IL15, GFP transposition was assessed. Figure 6 shows that SPB mRNA efficiently mediated transposition of the GFP transposon into T lymphocytes. Importantly, T cell viability was improved when co-delivering the SPB as an mRNA as opposed to a pDNA; 32.4% versus 25.4%, respectively. These data suggest that co-delivery of SPB as mRNA would be dose-sparing in the total amount of plasmid DNA being delivered to T cells and is thus less cytotoxic.

[0427] Since the current plasmid transposon also contains a backbone required for plasmid amplification in bacteria, it is possible to significantly reduce the total amount of DNA by excluding this sequence. This may be achieved by restriction digest of the plasmid transposon prior to the nucleofection reaction. In addition, this could be achieved by administering the transposon as a PCR product or as a doggybone™ DNA, which is a double stranded DNA that is produced in vitro by a mechanism that excludes the initial backbone elements required for bacterial replication of the plasmid.

[0428] We performed a pilot experiment to see whether or not plasmid transposon needed to be circular, or if it could be delivered to the cell in a linear fashion. To test this, transposon was incubated overnight with a restriction enzyme (ApaLI) to linearize the plasmid. Either uncut or linearized plasmid is electroporated into primary T lymphocytes. GFP expression was assessed 2 days later. Figure 7 shows that linearized plasmid was also efficiently delivered to the cell nucleus. These data demonstrate that linear transposon products can also be efficiently electroporated into primary human T cells.

[0429] We show above that plasmid DNA is toxic in primary T lymphocytes, but we have observed that this toxic effect is not as dramatic in tumor cell lines and other transformed cells. Based upon this observation, we hypothesized that primary T lymphocytes may be refractory to plasmid DNA transfection due to heightened DNA sensing pathways, which would protect immune cells from infection by viruses and bacteria. If these data are a result of heightened DNA sensing mechanisms, then it may be possible to enhance plasmid transfection efficiency and/or cell viability by the addition of DNA sensing pathway inhibitors to the post-nucleofection reaction. Thus, we tested a number of different reagents that inhibited the TLR-9 pathway, caspase pathway, or those involved in cytoplasmic double stranded DNA sensing. These reagents include Bafilomycin Al, which is an autophagy inhibitor that interferes with endosomal acidification and blocks NFkB signaling by TLR9, Chloroquine, which is a TLR9 antagonist, Quinacrine, which is a TLR9 antagonist and a cGAS antagonist, AC-YVAD-CMK, which is a caspase 1 inhibitor targeting the AIM2 pathway, Z-VAD-FMK, which is a pan caspase inhibitor, Z-IETD-FMK, which is a caspase 8 inhibitor triggered by the TLR9 pathway. In addition, we also tested the stimulation of electroporated T cells by the addition of the cytokines IL7 and IL15, as well as the addition of anti-CD3 anti-CD28 Dynabeads® Human T-Expander CD3/CD28 beads. Results are displayed in Figure 8. We found that few of the compounds or caspase inhibitors had any positive effect on cell viability at day 4 post-nucleofection at the doses tested. However, we acknowledge that further dosing studies may be required to better test these reagents. It may also be more effective to inhibit these pathways genetically. Two post-nucleofection conditions did enhance viability of the T cells. The addition of IL7 and IL15, whether they were added either 1 hour or 1 day following electroporation, enhanced viability over 3-fold when compared with introduction of the plasmid transposon alone without additional treatment. Furthermore, stimulation of the T cells post-nucleofection using either activator or expander beads also dramatically enhanced T cell viability; stimulation was better when the beads were added 1 hour or 1 day post-nucleofection as compared to adding the beads 2 days post. Lastly, we also tested ROCK inhibitor and the removal of dead cells from the culture using the Dead Cell Removal kit from Miltenyi, but saw no improvement in cell viability.

[0430] To further expand upon these findings demonstrating that stimulation of the T cells post-nucleofection improves viability, we repeated the study using the addition of the cytokine IL7 and IL15. Figure 9 shows that the addition of these cytokines each at a dose of 20 ng/mL either immediately following nucleofection or up to 1 hour post enhanced cell viability up to 2.9-fold when compared to no treatment. Addition of these cytokines up to 1 day post-nucleofection also enhanced viability, but not as strong as the prior time points.

[0431] Since we found that immediate stimulation of the T cells post-nucleofection was able to increase cell viability, we hypothesized that stimulating the cells prior to nucleofection may also enhance viability and transfection efficiency. To test this, we stimulated primary T lymphocytes either 2, 3, or 4 days prior to transposon nucleofection. Figure 10 shows that some level of transposition occurs when the transposon and the transposase are co-delivered after the T cells have been stimulated prior to the nucleofection reaction. The efficacy of pre- stimulation may be influenced by the kinetics of stimulation and may therefore be dependent upon the precise type of expander technology chosen.

EXAMPLE 2: Ex vivo genetic modification of NK cells

[0432] The piggyBac™ (PB) transposon system was used for genetically modifying human NK cells. Non-activated NK cells derived from CD3-depleted leukopheresis (containing CD14/CD19/CD56+ cells) were were electroporated with plasmid piggyBac transposon DNA encoding GFP and mRNA encoding Super piggyBac transposase using the program indicated in Figure 14 from Lonza 4D nucleofector or BTX ECM 830 (500V, 700 usee pulse length, 0.2 mm electrode gap, one pulse). Transposed cells were co-cultured (stimulated) at day 2 with artificial antigen presenting cells (aAPCs). Fluorescent activated cell sorting (FACS) analysis of GFP percent at day 7 post-EP (day 5 post-stimulation) is shown in Figure 14. Percent viability is the percentage of 7-Aminoactinomycin (7AAD)-negative cells at day 2 post-EP.

[0433] Transposition of non-activated NK cells from CD3-depleted leukopheresis

(containing CD14/CD19/CD56+ cells) is shown in Figure 15. Cells were electroporated with a plasmid piggyBac transposon encoding GFP and 5 ug mRNA encoding Super piggyBac transposase using the indicated Maxcyte electroporator program. Transposed cells were stimulated at day 2 with artificial antigen presenting cells (aAPCs). FACS plots (Figure 15 A) and a bar graph (Figure 15B) from the analysis of percent GFP+ of CD56+ cells at day 6 post-EP and day 4 post-stimulation are shown. Percent viability is the percentage of 7AAD- negative cells at day 2 post EP.

[0434] Figure 16 shows that there is dose-dependent DNA-mediated cytotoxicity in NK cells. FACS analysis of live cells (7AAD-ve/FSC, or Forward Scatter) at day 2 post-EP using Lonza 4D Nucleofector program DN-100. FACS plots (Figure 16A) are quantified in graph (Figure 16B). 5x10E6 cells were electroporated per electroporation in 100 uL P3 buffer in cuvettes. Cells were electroporated with no DNA (Mock) or varying amounts of piggyBac GFP transposon co-delivered with 5 ug super piggyBac mRNA.

EXAMPLE 3: In vitro differentiation of piggyBac modified HSPCs into B cells

[0435] Human CD34+ HSPCs were electroporated with mRNA encoding Super piggyBac along with a piggyBac transposon encoding GFP. After electroporation, HSPCs were primed for B cell differentiation in presence of human IL-3, Flt3L, TPO, SCF, and G-CSF for 5 days. On day 6, cells were transferred to a layer of MS-5 feeder cells and fed bi-weekly, along with transfer to a fresh layer of feeders once per week. On day 34 of the in vitro differentiation process, CD 19+ B cells were generated and detectable in the culture (Figure 17). A fraction of the B cells were positive for the GFP piggyBac transgene (Figure 17, lower right panel) demonstrating that the piggyBac DNA Modification System can be used to modify HSPCs, which can then be later differentiated into more differentiated immune cell types. This technique allows for the derivation of genetically-modified immune cells from hematopoietic progenitors.