TITLE OF THE INVENTION

Compositions and Methods Comprising Viral Vector Systems for Multiplexed Activation of Endogenous Genes as Immunotherapy and Viral -based Immune-Gene Therapy

CROSS-REFERENCE TO RELATED APPLICATION The present application is entitled to priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 62/924,973 filed October 23, 2019, which is hereby incorporated by reference in its entirety herein.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with government support under CA238295, CA231112, CA209992 and CA225498 awarded by the National Institutes of Health; and under W81XWH-17-1-0235 and W81XWH-20- 1-0072 awarded by the United States Army Medical Research and Material Command. The government has certain rights in the invention.

BACKGROUND OF THE INVENTION Immunotherapy has transformed cancer treatment by leveraging the patient’s own immune system against the tumor, thereby turning several previously lethal cancers into manageable diseases for a subset of patients. Major types of immunotherapy include checkpoint blockade, adoptive cell transfer, human recombinant cytokines, and cancer vaccines. Although antigen recognition is a key process in the anti-tumor immune response, there has been limited success using cancer vaccines in the clinic over the past several decades. Traditional cancer vaccines are often dendritic cell-based vaccines such as sipuleucel-T. Recent advances of peptide- and RNA-based vaccines showed that targeted delivery of multiple mutated neoantigens can generate a strong anti-tumor effect, providing direct clinical evidence of effective cancer vaccines against late-stage melanoma. Tumor cells harbor a multitude of mutations that frequently encode mutated, partially truncated, or amplified genes that are immunogenic. However, many of these mutations might not be expressed at levels sufficient to elicit an effective T-cell-mediated response. Synthesis of these peptides or transcripts is possible with parallel protein- or RNA-synthesis, but the cost of this approach is proportional to the number of mutations identified. A need exists for new types of cancer vaccines that can elicit strong, durable, and specific immune responses against cancer antigens, while maintaining efficacy, versatility, and cost-effectiveness. The present invention addresses this need.

SUMMARY OF THE INVENTION

As described herein, the present invention relates to compositions and methods comprising viral vector systems for multiplexed activation of endogenous genes as immunotherapy and viral -based immune-gene therapy.

In one aspect, the invention comprises a method of developing a cancer immunotherapy. The method comprises a) administering a CRISPR activation (CRISPRa) system comprising an sgRNA library to a cancer cell, thereby generating a modified cell, b) administering the modified cell to an immunocompetent mammal whereby the mammal develops cancer, c) determining the sgRNAs and thereby the targeted genes that are depleted in the cancer, and d) designing a cancer immunotherapy that targets the depleted genes.

In certain embodiments, the sgRNA library comprises the nucleotide sequences set forth in SEQ ID NOs. 86-192 or SEQ ID NOs. 193-411.

In certain embodiments, the mammal is a mouse.

In certain embodiments, the CRISPRa system comprises a vector comprising the nucleotide sequence set forth in SEQ ID NO: 1.

In certain embodiments, determining the genes that are depleted in the cancer comprises nucleotide sequencing and analysis.

In certain embodiments, designing a cancer therapy that targets the depleted genes comprises packaging the open reading frames (ORFs) of the depleted genes in a vector.

In certain embodiments, the vector is an adeno-associated viral (AAV) vector or an adenoviral vector.

In another aspect, the invention includes a method of treating cancer in a subject in need thereof. The method comprises administering a therapeutically effective amount of the cancer immunotherapy generated by any of the methods contemplated herein.

In another aspect, the invention includes a composition comprising a vector comprising an open reading frame (ORF) of at least one gene selected from the group consisting of CD80, Light, CXCL10, 4-1BBL, GITRL, IL2, IL-23, and IFNg.

In certain embodiments, the vector comprises the nucleotide sequence set forth in any one of SEQ ID NOs: 2, 3, and 24-58. In certain embodiments, the vector is an AAV vector or an adenoviral vector.

In another aspect, the invention includes a vector comprising a nucleic acid sequence that is 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NOs: 1-18 or 24-58.

In another aspect, the invention includes a vector comprising at least one nucleic acid sequence that is 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to any one of SEQ ID NOs: 73-84.

In certain embodiments, the vector is an AAV vector or an adenoviral vector.

In another aspect, the invention includes a composition comprising any of the vectors contemplated herein.

In another aspect, the invention includes a method of treating cancer in a subject in need therof. The method comprises administering to the subject a therapeutically effective amount of any of the compositions contemplated herein.

In certain embodiments, the cancer is selected from the group consisting of triple negative breast cancer, melanoma, pancreatic cancer, or a solid tumor.

In certain embodiments, the method further comprises administering an additional treatment to the subject.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description of specific embodiments of the invention will be better understood when read in conjunction with the appended drawings. For the purpose of illustrating the invention, there are shown in the drawings exemplary embodiments. It should be understood, however, that the invention is not limited to the precise arrangements and instrumentalities of the embodiments shown in the drawings.

Figs. 1 A-1D illustrate an in vivo CRISPRa screen to pinpoint the genetic factors that can enhance host immune surveillance and tumor destruction. Fig. 1 A is a schematic of the experiment design. The APCM sgRNA library was introduced into E0771, a triple negative breast cancer cell line, by infection with lentiviruses at M.O.I = 0.2-0.3. After 7 days of in vitro culture under puromycin selection, 2*10 ⁶ of APCM library -transduced cells were intravenously injected into immunocompetent C57BL/6J mice. The resulting metastatic tumors in the lungs were collected, and the immune-mediated sgRNA depletion and enrichment was readout by illumina sequencing. Fig. IB shows Kaplan- Meier survival curves of mice transplanted with E0771 -Vector (n = 10) or E0771-APCM library (n = 15) formed metastatic tumors. Log-rank test, E0771-APCM vs. E0771- Vector,/? = 0.2000. Fig. 1C depicts the correlations within tumor samples and cell samples. Fig. ID is a scatter plot of genes whose activation may lead to elevated immune destruction. Average sgRNA representation across metastatic tumors in C57BL/6J mice (n = 14) was plotted against average sgRNA library representation in cultured cell pool (n = 2). Screen was performed in immunocompetent C57BL/6J mice by intravenously injecting library -transduced E0771 to form metastatic tumors. The linearized sgRNA population regression was used to call depletion under immune pressure. Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001. Fig. IE depicts representative flow cytometry plots demonstrating CRISPRa- mediated expression of several representative costimulatory molecules, such as OX40L, ICOSL, CD80, CD86, and CD40 s in APCM sgRNA library transduced E0771 cells. Fig. IF shows the percentage of CD40, CD40L, CD80, CD86, OX40L, and ICOSL expressing cells in APCM sgRNA library transduced E0771 cells.

Fig. 2 illustrates in vivo efficacy of AAV-0-MAEGI-CLC4G, AAV-ORF-CLC4 and AAV-ORF-CLC4I in a syngeneic triple-negative breast cancer model. Therapeutic effects of intratumoral activation of CLC4I (Cd80, Iight/Tnfsfl4, CxcllO, 4- 1BBL/Tnfsf9, Ifng or Gitrl) are shown. Tumor growth curves of E0771 tumors treated with PBS (n = 6), AAV-dCas9 + Vector (n = 6), AAV-dCas9 + CLC4G (dual AAV delivered CRISPRa of sgRNA library targeting CLC4G, i.e. 0-MAEGI-CLC4G, n = 6), AAV-orf-CLC4 (pGW066: AAV delivered open reading frame expression of CLC4, n = 6), or AAV-orf-CLC4I (pGW067: AAV delivered open reading frame expression of Cleg and Ifng, n = 6). Two-way ANOVA: AAV-dCas9 + Vector vs. PBS, p = 0.9675; AAV- dCas9 + CLC4 vs. PBS,/? < 0.0001; AAV-orf-CLC4 vs. PBS,/? = 0.04; AAV-orf-CLC4I vs. PBS, p < 0.0001; AAV-dCas9 + CLC4G vs. AAV-dCas9 + Vector, p < 0.0001. Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05,

** p < 0.01, *** p < 0.001.

Figs. 3A-3C illustrate the design and characterization of an adenoviral CRISPRa system. Fig. 3 A is a schematic of the designs of adenoviral dSpCas9-based CRISPRa systems, that use a single construct to deliver all CRISPRa components. Depicted are pGW029 (SEQ ID NO: 4), pGW035 (SEQ ID NO: 5), and pGW063 (SEQ ID NO: 6).

Fig. 3B shows results from an experiment wherein 0x401 -targeted sgRNA was cloned into a construct from Fig. 3 A and the activation efficency tested in vitro. 4 days post- transfection, expression levels of 0x401 were determined by quantitative RT-PCR. Fig.

3C shows the cytopathic effects in HEK293FT cells caused by the adenoviral CRISPRa system. Upper panel: AdTrack (adenoviral CMV-EGFP) were rescued, and the expression of GFP and cytopathic effects were observed, demonstrating the robustness of the adenoviral engineering and rescue system. Lower panel: the CRISPRa system was cloned into adenoviral constructs (U6-sgRNA-EFS-LTR-dSpCas9-p65-HSFl-sPA) and the adenoviruses were rescued. 4-6 days post-infection of the adenovirus, cytopathic effects on HEK293FT were observed.

Figs. 4A-4B illustrate an AAV based one-vector system with dSaCas9. Fig. 4A is a diagram illustrating a dSaCas9-based AIO (all-in-one) CRISPRa system, using a single AAV vector to deliver all CRISPRa components. Fig. 4B shows in vitro activation of the Pmel gene with the AIO CRISPRa system. Pmel was targeted with the AIO CRISPRa system and 4 days post-infection expression levels of Pmel were determined by quantitative RT-PCR.

Figs. 5A-5B illustrate an AAV based two-vector system with dSaCas9. Fig 5A is a schematic of the design of a dSaCas9-based dual-AAV delivery CRISPRa system. Fig. 5B shows results from in vitro testing of the dSaCas9-based dual-AAV CRISPRa system. Pmel targeted sgRNA was cloned into with the dual AAV-CRISPRa system. Four days post-infection, expression levels of Pmel were determined by quantitative RT-PCR.

Figs. 6A-6B illustrate an AAV based two-vector system with dSpCas9. Fig. 6A is a schematic of the design of a dSpCas9-based dual-AAV delivery of CRISPRa system. Fig. 6B shows results of in vitro testing. Pmel targeted sgRNA was cloned into with the dual AAV-CRISPRa system. 6 days post-infection, expression levels of Pmel were determined by quantitative RT-PCR.

Fig. 7 is a table illustrating sgRNA sequences for an AAV-APCM library (SEQ ID NOs: 86-192).

Figs. 8A-8B illustrate sgRNA sequences for an AAV-APCM library designed/optimized specifically for human genes (SEQ ID NOs: 193-411).

Figs. 9A-9G illustrate multiplexed activation of endogenous genes as immunotherapy, by activating immune genes (APCM) or mutant genes (p-MAEGI). Fig. 9A shows tumor growth curves of orthotopic breast tumor formed by E0771-dCas9-VP64 in mice treated with PBS (n = 10 mice), AAV-Vector (n = 10), or AAV-APCM (n = 8) by intratumoral administration at indicated times (arrows). Two-way ANOVA: AAV-Vector vs. PBS, ? = 0.0054; AAV- APCM vs. PBS,/? < 0.0001; AAV- APCM vs. AAV-Vector, p = 0.0139. Fig. 9B illustrates the therapeutic effects of dual-AAV delivered APCM CRISPRa into tumor-bearing mice. The growth curves of E0771 syngeneic tumors in mice treated by intratumoral injection of PBS (n = 5 mice), AAV-dCas9 + Vector (n = 5), or AAV-dCas9 + APCM (n = 5) at the indicated time points (arrows). Two-way ANOVA: AAV-dCas9 + Vector vs. PBS, p = 0.1586; AAV-dCas9 + APCM vs. PBS, p < 0.0001; AAV-dCas9 + APCM vs. AAV-dCas9 + Vector, p < 0.0001. Fig. 9C shows an experimental design for evaluating the systemic anti -tumor effects of AAV-APCM. 2*10 ⁶ or 0.2* 10 ⁶ E0771-dCas9-VP64 tumor cells were transplanted into the left or right flank of C57BL/6J mice respectively to model local and distant tumors. AAV-p-MAEGI was administered only into the local tumors at the indicated times (arrows). Figs. 9D-9E are growth curves of E0771-dCas9-VP64 local (Fig. 9D) and distant (Fig. 9E) tumors in mice treated with PBS (n = 11), AAV-Vector (n = 11), or AAV-APCM (n = 17). Fig. 9D:

Local tumors, two-way ANOVA: AAV-Vector vs. PBS,/ _? = 0.0001; AAV- APCM vs. PBS,/ _? < 0.0001; AAV- APCM vs. AAV-Vector, = 0.0130. Fig. 9E: Distant tumor: two-way ANOVA: AAV-Vector vs. PBS,/? = 0.2308; AAV- APCM vs. PBS,/? = 0.011; AAV- APCM vs. AAV-Vector,/ _? = 0.1407. Fig. 9F shows survival curves of the E0771 breast tumor bearing mice treated with PBS (n = 6), AAV-Vector (n = 13), or AAV- APCM (n = 12). Log-rank test, AAV-Vector vs. PBS,/ _? = 0.1115; AAV- APCM vs. PBS, p = 0.0022; AAV- APCM vs. AAV-Vector,/ _? = 0.1178. Fig. 9G show growth curves of Pan02-dCas9-VP64 tumors in C57BL/6J mice treated with PBS (n = 6 mice), AAV- Vector (n = 6), AAV-APCM (n =7), or AAV-p-MAEGI (n = 8) at indicated times (blue arrows). Two-way ANOVA: AAV-Vector vs. PBS,/ _? < 0.0001; AAV- APCM vs. PBS,/ _? < 0.0001; AAV-p-MAEGI vs. PBS,/ _? < 0.0001; AAV- APCM vs. AAV- Vector, p = 0.4432; AAV-p-MAEGI vs. AAV-Vector, p < 0.0001. Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001.

Figs. 10A-10F illustrate therapeutic effects by multiplexed activation of Cd80, Light, CxcllO, Gitrl, 41bbl, and IFNg in tumor cells by AAV-delivered CRISPRa or AAV-ORFs (polycistronic open-reading-frames). Fig. 10A left: AAV-CRISPRa-mediated transcriptional activation of Cd80, Light, CxcllO, Gitrl, 41bbl, and IFNg, normalized to vector-transduced controls (n = 8, mutiple t-test, AAV-sgRNAs versus vector Cd80, P= 0.01; Light, P = 0.00001; CxcllO, P = 0.0003; Gitrl, P = 0.0004; 41bbl, P = 0.00002; Ifny, P < 0.0001); Fig. 10A right: Co-transfection of Dual AAV-CRISPRa-mediated transcriptional activation of Cd80, Light, CxcllO, Gitrl, 41bbl, and IFNg, normalized to control (n = 4). Fig. 10B shows the expression of CD80, 41BBL, and Gitrl in dual-AAV- CRISPRa infected cells as assessed by flow cytometry. Fig. IOC shows the therapeutic effects of intratumoral injected AAV-CRISPRa. Dual-AAV mediated activation of Cd80, Iight/Tnfsfl4, CxcllO, 4-1 BBL/TnfsfO, and Ifng (CLC4I). Growth curves of E0771 tumors treated with PBS (n = 6), AAV-dCas9 + Vector (n = 6), AAV-dCas9 + CLCG4 (dual AAV delivered CRISRPa of Cleg (n = 6). Two-way ANOVA: AAV-dCas9 + Vector vs. PBS,/? = 0.9675; AAV-dCas9 + Clcg4 vs. PBS,/? < 0.0001; AAV-dCas9 + Clcg4 vs. AAV-dCas9 + Vector, p = 0.0028. Fig. 10D illustrates express Cd80, Light, CxcllO, 41bbl, or/and IFNg using AAV polycistronic ORFs. Both transfection and AAV infection lead to high levels’ expression of these molecules. Fig. 10E illustrates the therapeutic effects of intratumoral injected AAV-ORFs expressing Cd80, Light, CxcllO, 41bbl (orfClc4, n = 6), or AAV expressing Cd80, Light, CxcllO, 41bbl, Ifng (orfClc4I, n = 6). Fig. 10F illustrates the therapeutic effects of intratumoral injected AAV-ORFs expressing Cd80, Light, CxcllO, 41bbl, Ifng (orfClc4I, n = 5). Two-way ANOVA: AAV- orfCLC4 vs. PBS,/ _? < 0.0001; AAV- Vector vs. PBS, p = 0.0041; AAV-orfCLC4 vs. AAV- Vector, p = 0.0038. Two-way ANOVA: AAV-orfCLC4 vs. PBS,/ _? = 0.6182; AAV-orfCLC4I vs. PBS, p < 0.0001. Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001.

Figs. 11 A-l IF illustrate the finding that AAV-mediated costimulatory molecule expression promotes tumor immune infiltration. Fig. 11 A shows flow cytometry quantification of CD45+ immune cells in tumor microenvironment at DPI = 30. Percentage of CD45+ immune cells out of total cells in tumors treated with PBS, dual- AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I (n = 5 mice per group; two-tailed Mann-Whitney test: AAV-dCas9 + Clcg4 vs. PBS, P = 0.0931; AAV-orfClc4I vs. PBS, P = 0.0411). Fig. 1 IB shows percentage of CD8+ T cells out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV- orfClc4I (n = 5 mice per group; two-tailed Mann-Whitney test: AAV-dCas9 + Clcg4 vs. PBS, P = 0.0411; AAV-orfClc4I vs. PBS, P = 0.0411). Fig. 11C shows percentage of CD4+ T cells out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV- Clcg4, AAV-orfClc4, AAV-orfClc4I (n = 5 mice per group; two-tailed Mann-Whitney test: AAV-dCas9 + Clcg4 vs. PBS, P = 0.1320; AAV-orfClc4I vs. PBS, P = 0.0152). Fig.

1 ID shows flow cytometry quantification of PD1+CD8+ T cells out of total CD8+ T cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV- orfClc4I (n = 5 mice per group; two-tailed Mann-Whitney test: AAV-dCas9 + Clcg4 vs. PBS, P = 0.1797; AAV-orfClc4I vs. PBS, P = 0.026). Fig. 1 IE shows flow cytometry quantification of monocytes out of total cells in tumors treated with PBS, dual-AAV- vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I (n = 5 mice per group; two-tailed Mann-Whitney test: AAV-dCas9 + Clcg4 vs. PBS, P = 0.0411; AAV-orfClc4I vs. PBS, P = 0.6991). Fig. 1 IF shows flow cytometry quantification of dendritic cells out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I (n = 5 mice per group; two-tailed Mann-Whitney test: AAV-dCas9 + Clcg4 vs. PBS, P = 0.0411; AAV-orfClc4I vs. PBS, P = 0.5887). Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001.

Figs. 12A-12F illustrate optimized combinations of immune-stimulating molecules for tumor therapy. Fig. 12A illustrates the therapeutic effects of intra-tumoral injected AAV-CRISPR activating Cd80, light, CxcllO, 4-1BBL, Gitrl and Ifng (Clcg4I). Growth curves of E0771 tumors treated with PBS (n = 5), AAV-Vector (pGW045; n = 5), or AAV-Clcg4I (pGW045-Clcg4I). Two-way ANOVA: AAV- Vector vs. PBS, p = 0.0871; AAV-Clcg4 vs. PBS,/? < 0.0001; AAV- Clcg4 vs. AAV-Vector, p = 0.0009. Figs. 12B-12C illustrate further optimization of the combinatorial genes pool by removing one gene in each pool. Fig. 12B shows growth curves of E0771 tumors treated with PBS (n = 4), AAV-Vector (n = 4), AAV-Clcg4I (n = 4), or AAV-Clcg4Iminusl (n =

4). Two-way ANOVA: AAV-Vector vs. PBS,/? = 0.0002; AAV- Clcg4I vs. AAV- Vector, p = 0.0002. Fig. 12C shows growth curves of E0771 tumors treated with AAV- Clcg4I (n = 4), or AAV-Clcg4I minusl (n = 4). Two-way ANOVA: AAV- Clcg4I vs. 41BBL, p < 0.0001; AAV- Clcg4I vs. -IFNg, p = 0.05; AAV- Clcg4I vs. -CD80, p = 0.095; AAV- Clcg4I vs. -Light, p = 0.05; AAV- Clcg4I vs. -Gitrl, p = 0.0116. Fig. 12D shows growth curves of B16F10 melanoma treated with PBS (n = 4), AAV-Vector (n =

5), AAV-p67:Clc4I (Cd80, Light, CxcllO, 41bbl, Ifng; n = 5), AAV-P72:IFNg (n = 4). Two-way ANOVA: AAV-Vector vs. PBS,/? = 0.3495; AAV-p67:Clc4I vs. Vector,/? = 0.0012; AAV-p72:IFNg vs. PBS, p < 0.0001. Fig. 12E shows growth curves of E0771 tumors treated with PBS (n = 6), AAV-Vector (n = 6), AAV-P72:IFNg (n = 6), AAV- p94:IClG4 (Ifng, Cd80, Light, Gitrl, 41bbl; n = 6). Fig. 12F shows growth curves of E0771 tumors treated with AAV-Vector (n = 5), AAV-polycistronic PI 19:IFNg-STOP- CD80-Light-Cxcl 10-41BBL (n = 5), AAV-polycistronic P 118 :IFNg-CD80-Light-Cxcl 10- 41BBL (n = 5), AAV-polycistronic P100:IFNg-CD80-Light-Cxcll0 (n = 5), AAV-P67: CD80-Light-Cxcl 10-4 lBBL-IFNg (n = 5). Two-way ANOVA: AAV- Vector vs. PBS, p = 0.0455; AAV-p94:ICLG4 vs. Vector,/? = 0.0111; AAV-p72:IFNg vs. PBS, p < 0.0001. Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05,

** p < 0.01, *** p < 0.001

Figs. 13A-13B illustrate the expression levels of IFNg and CD80 in transfected cells using different polycistronic AAV-ORFs. Fig. 13A Upper panel: Representative flow cytometry plots demonstrating AAV-ORFs-mediated expression of IFNg in different AAV constructs transfected E0771 cells; low panel (left): the percentage of IFNg expressing cells in different constructs-transfected E0771 cells; low panel (right): the median fluorescent intensity (MFI) of IFNg-APC in different constructs-transfected E0771 cells. Fig. 13B Upper panel: Representative flow cytometry plots demonstrating AAV-ORFs-mediated expression of CD80 in different AAV constructs transfected E0771 cells; low panel (left): the percentage of CD80+ cells in different constructs-transfected E0771 cells; low panel (right): the median fluorescent intensity (MFI) of CD80-PE in different constructs-transfected E0771 cells.

Figs. 14A-14F illustrate optimizing costimulatory molecule combinations for tumor therapy. Fig. 14A Upper panel: Representative flow cytometry plots demonstrating AAV-ORFs-mediated expression of CD80 and 41BBL in AAV-infected E0771 or MA1NC cells; lower panel: the percentage of CD80 and 41BBL positive cells in different AAV-infected E0771 and MA1NC cells. Fig. 14B Upper panel: Representative flow cytometry plots demonstrating AAV-ORFs-mediated expression of IL23 and IFNg in AAV-infected E0771 or MA1NC cells; lower panel: the percentage of IL23 and IFNg positive cells in different AAV-infected E0771 and MA1NC cells. Fig. 14C shows tumor growth curves of E0771 tumors treated with PBS (n = 5), AAV-Vector (n = 10), AAV- IL23 (n = 5), AAV-IFNg (n = 6), Pooled AAVs(IFNg + CD80 + LIGHT + CXCL10 + 41BBL; n = 5). Fig. 14D shows tumor growth curves of E0771 tumors treated AAV- Vector (n =5), AAV-IFNg (n = 4), AAV-IFNg+AAV-IL23 (n = 5), pooled AAVs (IFNg + CXCL10 + 41BBL; n = 5), pooled AAVs (IFNg + LIGHT + 41BBL; n = 5), pooled AAV(IFNg + LIGHT + CXCL10 + 41BBL; n = 5). Fig. 14E shows tumor growth curves of E0771 tumors treated AAV-Vector (n =11), AAV-IFNg (n = 10), AAV-IL23 (n = 10), AAV-IFNg+AAV-IL23 (n = 4). Fig. 14F shows tumor growth curves of E0771 tumors treated AAV-Vector (n = 5), AAV-IFNg (n = 5), AAV-IL23 (n = 5), AAV-IFNg+AAV- IL23+AAV-41BBL (n = 5). Error bars: Data points in this figure are presented as mean ± s.e.m. Asterisks: * p < 0.05, ** p < 0.01, *** p < 0.001.

Fig. 15 depicts schematics of the construct designs for AAV-ORFs. Constructs include: pGW066 inserts(including ITR): 4542 bp (SEQ ID NO: 2); pGW067 inserts(including ITR): 4513 bp (SEQ ID NO: 3); pGW071 inserts(including ITR): 4556 bp (SEQ ID NO: 24); pGW072 inserts(including ITR): 1360 bp (SEQ ID NO: 25); pGW073 inserts(including ITR): 4567 bp (SEQ ID NO: 26); pGW078 inserts(including ITR): 895 bp (SEQ ID NO: 27); pGW071 inserts(including ITR): 4556 bp (SEQ ID NO: 24); pGW089 inserts(including ITR): 1401 bp (SEQ ID NO: 57); pGW090 inserts(including ITR): 4726 bp (SEQ ID NO: 28); pGW091 inserts(including ITR): 4738 bp (SEQ ID NO: 29); pGW094 inserts(including ITR): 4723 bp (SEQ ID NO: 58); pGW096 inserts(including ITR): 5209 bp (SEQ ID NO: 30); pGW097 inserts(including ITR): 1899 bp (SEQ ID NO: 31); pGW098 inserts(including ITR): 4207 bp (SEQ ID NO: 32); pGW099 inserts(including ITR): 3496 bp (SEQ ID NO: 33); pGWIOO inserts(including ITR): 4498 bp (SEQ ID NO: 34); pGWIOl inserts(including ITR): 4132 bp (SEQ ID NO: 35); pGW102 inserts(including ITR): 1816 bp (SEQ ID NO: 36); pGW103 inserts(including ITR): 1612 bp (SEQ ID NO: 37); pGW104 inserts(including ITR): 1189 bp (SEQ ID NO: 38); pGW105 inserts(including ITR): 1822 bp (SEQ ID NO: 39); pGWl 10 inserts(including ITR): 4080 bp (SEQ ID NO: 40); pGWl 11 inserts(including ITR): 3093 bp (SEQ ID NO: 41); pGW112 inserts(including ITR): 4083 bp (SEQ ID NO: 42); pGWl 13 inserts(including ITR): 4161 bp (SEQ ID NO: 43); pGW114 inserts(including ITR): 4713 bp (SEQ ID NO: 44); pGW115 inserts(including ITR): 4794 bp (SEQ ID NO: 45); pGWl 18 inserts(including ITR): 4133 bp (SEQ ID NO: 46); pGWl 19 inserts(including ITR): 4133 bp (SEQ ID NO: 47); pGW122 inserts(including ITR): 4002 bp (SEQ ID NO: 48); pGW127b inserts(including ITR): 3852 bp (SEQ ID NO: 49); pGW128b inserts(including ITR): 4671 bp (SEQ ID NO: 50); pGW145 inserts(including ITR): 2023 bp (SEQ ID NO: 51); pGW146 inserts(including ITR): 2509 bp (SEQ ID NO: 52); pGW147 inserts(including ITR): 2287 bp (SEQ ID NO: 53); pGW149 inserts(including ITR): 2260 bp (SEQ ID NO: 54)

DETAILED DESCRIPTION

Definitions Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present invention, the preferred materials and methods are described herein. In describing and claiming the present invention, the following terminology will be used.

It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

“About” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of ±20% or ±10%, more preferably ±5%, even more preferably ±1%, and still more preferably ±0.1% from the

As used herein, the term “autologous” is meant to refer to any material derived from the same individual to which it is later to be re-introduced into the individual.

“Allogeneic” refers to any material derived from a different animal of the same species.

As used herein, the term “bp” refers to base pair.

The term “complementary” refers to the degree of anti-parallel alignment between two nucleic acid strands. Complete complementarity requires that each nucleotide be across from its opposite. No complementarity requires that each nucleotide is not across from its opposite. The degree of complementarity determines the stability of the sequences to be together or anneal/hybridize. Furthermore, various DNA repair functions as well as regulatory functions are based on base pair complementarity.

The term “CRISPR/Cas” or “clustered regularly interspaced short palindromic repeats” or “CRISPR” refers to DNA loci containing short repetitions of base sequences followed by short segments of spacer DNA from previous exposures to a virus or plasmid. Bacteria and archaea have evolved adaptive immune defenses termed CRISPR/CRISPR-associated (Cas) systems that use short RNA to direct degradation of foreign nucleic acids. In bacteria, the CRISPR system provides acquired immunity against invading foreign DNA via RNA-guided DNA cleavage.

The “CRISPR/Cas9” system or “CRISPR/Cas9-mediated gene editing” refers to a type II CRISPR/Cas system that has been modified for genome editing/engineering. It is typically comprised of a “guide” RNA (gRNA) and a non-specific CRISPR-associated endonuclease (Cas9). “Guide RNA (gRNA)” is used interchangeably herein with “short guide RNA (sgRNA)” or “single guide RNA (sgRNA). The sgRNA is a short synthetic RNA composed of a “scaffold” sequence necessary for Cas9-binding and a user-defined ~20 nucleotide “spacer” or “targeting” sequence which defines the genomic target to be modified. The genomic target of Cas9 can be changed by changing the targeting sequence present in the sgRNA.

“CRISPRa” system refers to a modification of the CRISPR-Cas9 system that functions to activate or increase gene expression. In certain embodiments, the CRISPRa system is comprised of a catalytically dead RNA/DNA guided endonuclease, such as dCas9, dCasl2a/dCpfl, dCasl2b/dC2cl, dCasl2c/dC2c3, dCasl2d/dCasY, dCasl2e/dCasX, dCasl3a/dC2c2, dCasl3b, dCasl3c, dCasl4, dead Cascade complex, or others; at least one transcriptional activator; and at least one sgRNA that functions to increase expression of at least one gene of interest. The term “activation” as used herein refers to an increase in gene expression of one or more genes.

“dCas9” as used herein refers to a catalytically dead Cas9 protein that lacks endonuclease activity. “dSaCas9” refers to dCas9 derived from Staphylococcus aureus. dSpCas9” refers to dCas9 derived from Streptococcus pyogenes.

A “disease” is a state of health of an animal wherein the animal cannot maintain homeostasis, and wherein if the disease is not ameliorated then the animal’s health continues to deteriorate. In contrast, a “disorder” in an animal is a state of health in which the animal is able to maintain homeostasis, but in which the animal’s state of health is less favorable than it would be in the absence of the disorder. Left untreated, a disorder does not necessarily cause a further decrease in the animal’s state of health.

The term “downregulation” as used herein refers to the decrease or elimination of gene expression of one or more genes.

“Effective amount” or “therapeutically effective amount” are used interchangeably herein, and refer to an amount of a compound, formulation, material, or composition, as described herein effective to achieve a particular biological result or provides a therapeutic or prophylactic benefit. Such results may include, but are not limited to, anti-tumor activity as determined by any means suitable in the art.

“Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.

As used herein “endogenous” refers to any material from or produced inside an organism, cell, tissue or system.

As used herein, the term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue or system.

The term “expression” as used herein is defined as the transcription and/or translation of a particular nucleotide sequence driven by its promoter.

“Expression vector” refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids ( e.g ., naked or contained in liposomes) and viruses ( e.g ., Sendai viruses, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.

“Homologous” as used herein, refers to the subunit sequence identity between two polymeric molecules, e.g., between two nucleic acid molecules, such as, two DNA molecules or two RNA molecules, or between two polypeptide molecules. When a subunit position in both of the two molecules is occupied by the same monomeric subunit; e.g, if a position in each of two DNA molecules is occupied by adenine, then they are homologous at that position. The homology between two sequences is a direct function of the number of matching or homologous positions; e.g, if half (e.g, five positions in a polymer ten subunits in length) of the positions in two sequences are homologous, the two sequences are 50% homologous; if 90% of the positions (e.g, 9 of 10), are matched or homologous, the two sequences are 90% homologous.

“Identity” as used herein refers to the subunit sequence identity between two polymeric molecules particularly between two amino acid molecules, such as, between two polypeptide molecules. When two amino acid sequences have the same residues at the same positions; e.g, if a position in each of two polypeptide molecules is occupied by an Arginine, then they are identical at that position. The identity or extent to which two amino acid sequences have the same residues at the same positions in an alignment is often expressed as a percentage. The identity between two amino acid sequences is a direct function of the number of matching or identical positions; e.g. , if half (e.g, five positions in a polymer ten amino acids in length) of the positions in two sequences are identical, the two sequences are 50% identical; if 90% of the positions (e.g, 9 of 10), are matched or identical, the two amino acids sequences are 90% identical.

As used herein, an “instructional material” includes a publication, a recording, a diagram, or any other medium of expression which can be used to communicate the usefulness of the compositions and methods of the invention. The instructional material of the kit of the invention may, for example, be affixed to a container which contains the nucleic acid, peptide, and/or composition of the invention or be shipped together with a container which contains the nucleic acid, peptide, and/or composition. Alternatively, the instructional material may be shipped separately from the container with the intention that the instructional material and the compound be used cooperatively by the recipient.

“Isolated” means altered or removed from the natural state. For example, a nucleic acid or a peptide naturally present in a living animal is not “isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is “isolated.” An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.

The term “knockdown” as used herein refers to a decrease in gene expression of one or more genes.

The term “knockout” as used herein refers to the ablation of gene expression of one or more genes.

A “lentivirus” as used herein refers to a genus of the Retroviridae family. Lentiviruses are unique among the retroviruses in being able to infect non-dividing cells; they can deliver a significant amount of genetic information into the DNA of the host cell, so they are one of the most efficient vectors for gene delivery. HIV, SIV, and FIV are all examples of lentiviruses. Vectors derived from lentiviruses offer the means to achieve significant levels of gene transfer in vivo.

By the term “modified” as used herein, is meant a changed state or structure of a molecule or cell of the invention. Molecules may be modified in many ways, including chemically, structurally, and functionally. Cells may be modified through the introduction of nucleic acids.

By the term “modulating,” as used herein, is meant mediating a detectable increase or decrease in the level of a response in a subject compared with the level of a response in the subject in the absence of a treatment or compound, and/or compared with the level of a response in an otherwise identical but untreated subject. The term encompasses perturbing and/or affecting a native signal or response thereby mediating a beneficial therapeutic response in a subject, preferably, a human.

A “mutation” as used herein is a change in a DNA sequence resulting in an alteration from a given reference sequence (which may be, for example, an earlier collected DNA sample from the same subject). The mutation can comprise deletion and/or insertion and/or duplication and/or substitution of at least one deoxyribonucleic acid base such as a purine (adenine and/or thymine) and/or a pyrimidine (guanine and/or cytosine). Mutations may or may not produce discernible changes in the observable characteristics (phenotype) of an organism (subject).

By “nucleic acid” is meant any nucleic acid, whether composed of deoxyribonucleosides or ribonucleosides, and whether composed of phosphodiester linkages or modified linkages such as phosphotriester, phosphoramidate, siloxane, carbonate, carboxymethylester, acetamidate, carbamate, thioether, bridged phosphoramidate, bridged methylene phosphonate, phosphorothioate, methylphosphonate, phosphorodithioate, bridged phosphorothioate or sulfone linkages, and combinations of such linkages. The term nucleic acid also specifically includes nucleic acids composed of bases other than the five biologically occurring bases (adenine, guanine, thymine, cytosine and uracil). In the context of the present invention, the following abbreviations for the commonly occurring nucleic acid bases are used. “A” refers to adenosine, “C” refers to cytosine, “G” refers to guanosine, “T” refers to thymidine, and “U” refers to uridine.

Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain an intron(s).

The term “oligonucleotide” typically refers to short polynucleotides, generally no greater than about 60 nucleotides. It will be understood that when a nucleotide sequence is represented by a DNA sequence {i.e., A, T, G, C), this also includes an RNA sequence i.e., A, U, G, C) in which “U” replaces “T”.

As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

“Parenteral” administration of an immunogenic composition includes, e.g ., subcutaneous (s.c.), intravenous (i.v.), intramuscular (i.m.), or intrastemal injection, or infusion techniques.

As used herein, the terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof. The term “polynucleotide” as used herein is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Thus, nucleic acids and polynucleotides as used herein are interchangeable. One skilled in the art has the general knowledge that nucleic acids are polynucleotides, which can be hydrolyzed into the monomeric “nucleotides.” The monomeric nucleotides can be hydrolyzed into nucleosides. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any means available in the art, including, without limitation, recombinant means, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using ordinary cloning technology and PCR™, and the like, and by synthetic means. Conventional notation is used herein to describe polynucleotide sequences: the left-hand end of a single-stranded polynucleotide sequence is the 5’- end; the left-hand direction of a double-stranded polynucleotide sequence is referred to as the 5’ -directi on.

A “sample” or “biological sample” as used herein means a biological material from a subject, including but is not limited to organ, tissue, exosome, blood, plasma, saliva, urine and other body fluid. A sample can be any source of material obtained from a subject.

The term “subject” is intended to include living organisms in which an immune response can be elicited (e.g., mammals). A “subject” or “patient,” as used therein, may be a human or non-human mammal. Non-human mammals include, for example, livestock and pets, such as ovine, bovine, porcine, canine, feline and murine mammals. Preferably, the subject is human.

As used herein, a “substantially purified” cell is a cell that is essentially free of other cell types. A substantially purified cell also refers to a cell which has been separated from other cell types with which it is normally associated in its naturally occurring state. In some instances, a population of substantially purified cells refers to a homogenous population of cells. In other instances, this term refers simply to cell that have been separated from the cells with which they are naturally associated in their natural state. In some embodiments, the cells are cultured in vitro. In other embodiments, the cells are not cultured in vitro.

A “target site” or “target sequence” refers to a genomic nucleic acid sequence that defines a portion of a nucleic acid to which a binding molecule may specifically bind under conditions sufficient for binding to occur. The term “therapeutic” as used herein means a treatment and/or prophylaxis. A therapeutic effect is obtained by suppression, remission, or eradication of a disease state.

The term “transfected” or “transformed” or “transduced” as used herein refers to a process by which exogenous nucleic acid is transferred or introduced into the host cell. A “transfected” or “transformed” or “transduced” cell is one which has been transfected, transformed or transduced with exogenous nucleic acid. The cell includes the primary subject cell and its progeny.

To “treat” a disease as the term is used herein, means to reduce the frequency or severity of at least one sign or symptom of a disease or disorder experienced by a subject.

As used herein, “vaccinating” means administering a substance to a subject that induces an immune response against a disease.

A “vector” is a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to include non-plasmid and non- viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, and the like. Examples of viral vectors include, but are not limited to, Sendai viral vectors, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

Ranges: throughout this disclosure, various aspects of the invention can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

Description The present invention includes compositions and methods for treating cancer. In certain embodiments, the invention includes methods for treating cancer using a viral vector based (such as Adenovirus or AAV-based ) composition that utilizes the CRISPR activation (CRISPRa) system to activate endogenous genes. In certain embodiments, the invention includes use of the CRISPRa system to identify key factors for generating direct ORF-based immune-gene therapies.

It was discovered herein that direct activation of endogenous genes using a single vector system is an effective means to amplify tumor-associated antigens to enhance anti tumor immune responses. Highly multiplexed and customizable endogenous neoantigen activation using a single vector system has not yet been demonstrated prior to this disclosure. The present invention demonstrates the feasibility of using CRISPR technologies with RNA-guided precise genetic manipulations. The CRISPR activation (CRISPRa) system, based on dead Cas9 without nuclease activity (dCas9) (Qi et al.( 20131) Cell 152, 1173-1183), enables simple and flexible regulation of gene expression with dCas9 - transcriptional activation domain fusion (Gilbert et al. (2013) Cell 154, 442-451), which is further augmented by the recruitment of synergistic activation mediators (SAM) (Konermann, et al. (2015) Nature 517, 583-588; Chavez, et al. (2015) Nat Methods 12, 326-328; Tanenbaum, et al. (2014) Cell 159, 635-646).

Herein, the CRISPRa system was harnessed to manipulate endogenous gene expression to magnify anti-tumor immune responses. The approach was developed into multiplexed tumor vaccination strategies.

Another aspect of the invention includes compositions and methods comprising off-the-shelf endogenous gene vaccination cancer vaccines. The off-the-shelf versions have fixed components, which can have multiple forms, including single component or multi-component vaccines. The compositions of these forms of cancer vaccines and their pre-clinical efficacy showed that they can be an effective means of prophylactic and therapeutic agents.

CRISPR activation (CRISPRa) system

The CRISPR activation (CRISPRa) system is comprised of a catalytically inactive RNA-guided endonuclease or other endonucleases, such as but not limited to dCas9, dCasl2a/dCpfl, dCasl2b/dC2cl, dCasl2c/dC2c3, dCasl2d/dCasY, dCasl2e/dCasX, dCasl3a/dC2c2, dCasl3b, dCasl3c, dCasl4, dead Cascade complex, or others. The CRISPRa system also comprises at least one transcriptional activator, and at least one sgRNA that functions to increase expression of at least one gene of interest. Like a standard CRISPR-Cas9 system, CRISPRa systems rely on sgRNAs to guide Cas9 to intended targets. However, while a standard CRISPR-Cas9 system creates breaks in DNA through the endonuclease activity of Cas9 and then manipulates DNA repair mechanisms for gene editing, CRISPRa systems are modified and employ transcriptional activators to increase expression of genes of interest.

“dCas9” refers to a catalytically dead Cas9 protein that lacks endonuclease activity. This can be accomplished by introducing point mutations in the two catalytic residues (D10A and H840A) of the gene encoding Cas9. In doing so, dCas9 is unable to cleave dsDNA but retains the ability to target and bind DNA. This alone is often enough to attenuate if not outright block transcription of the targeted gene if the gRNA positions dCas9 in a way that prevents transcriptional factors and RNA polymerase from accessing the DNA. However, this ability to bind DNA can also be exploited for activation since dCas9 has modifiable regions, typically the N and C terminus of the protein, that can be used to attach transcriptional activators. dCas9 can be derived, for example, from S. pyogenes (dSpCas9), S. aureus (dSaCas9) N. meningiditis , S. thermopilus , F. novicida, C. jejuni, B. laterosporus, or from other species.

Targeting specificity of the CRISPRa system is determined by complementary base-pairing of a small guide RNA (sgRNA) to the genomic loci. sgRNA is a chimeric noncoding RNA that can be subdivided into three regions: a base-pairing sequence, a dCas9-binding hairpin and a terminator. When designing a synthetic sgRNA, only the base-pairing sequence is modified. Secondary variables must also be considered: off- target effects (for which a simple BLAST run of the base-pairing sequence is required), maintenance of the dCas9-binding hairpin structure, and ensuring that no restriction sites are present in the modified sgRNA, as this may pose a problem in downstream cloning steps. Due to the simplicity of sgRNA design, this technology is amenable to genome wide scaling.

Transcriptional activators are protein domains or whole proteins that can be linked to dCas9 or sgRNAs and assist in the recruitment of important co-factors as well as RNA Polymerase for transcription of the gene(s) targeted by the system. Transcriptional activators have a DNA binding domain and a domain for activation of transcription. The activation domain can recruit general transcription factors or RNA polymerase to the gene sequence. Activation domains can also function by facilitating transcription by stalled RNA polymerases, and in eukaryotes can act to move nucleosomes on the DNA or modify histones to increase gene expression. These activators can be introduced into the system through attachment to dCas9 or to the sgRNA. Transcriptional activators can be either mammalian cellular endogenous proteins that have activator function, activators from other species such as viruses, microbials or plants, their partial or mutant variants, engineered activators, or other forms of activators that can increase gene expression. A list of applicable viral activators include but are not limited to: VP 16, VP32, VP64, VP160, HBx, NS proteins, and VMW65. A list of applicable microbial activators include but are not limited to: Lac operons and GAL4. A list of applicable mammalian cellular transcriptional activators include but are not limited to: CAP, ACTN1, ACTN2, ACTN2, ACTN4, ACTN4, ANKRD1, APEX1, ARID5B, ARL2BP, ASCC1, ASXL1, ATN1, ATXN7L3, ATXN7L3, ATXN7L3, BCL9, BCL9L, BCL10, BCL10, BICRA, BIRC2, BRCA1, BRD7, CALCOCOl, CALCOCOl, CALCOCOl, CALCOCOl, CARM1, CARMl, CARM1, CBFB, CCARl, CCARl, CCARl, CCARl, CCAR2, CCDC62, CEBPA, CENPJ, CITED 1, CITED 1, CITED2, CITED2, CITED2, CITED2, CITED2, CITED4, CITED4, CITED4, COPS5, CREBBP, CREBBP, CREBBP, CTBP2, CTNNB1, CTNNB1, CTNNB1, CTNNB1, CTNNB1, DAXX, DAXX, DCAF6, DCC, DDX17, DHX9, DR1, DYRK1B, EDF1, ELF 3, ELOB, ENY2, ENY2, ENY2, EP300, EP300, EP300, FAM129B, FGF2, FHL5, FOXC1, GAT A3, GAT A3, GAT A3, GATA4, GM20517, GMEB1, GMEB2, GPS2, GPS2, GTF2A2, GTF2A2, FLAND1, HCFC1, HCFC1, HELZ2, HIF3A, HINFP, HIPK2, HMGA1, HMGA1, HMGA1B, HMGB2, HYAL2, ING4, ISL1, JADEl, JMJD6, JMY, JMY, JUN, JUN, JUNB, JUND, JUP, JUP, KAT2A, KAT2B, KAT2B, KAT5, KAT5, KAT6A, KDM1A, KDM5A, KMT2C, KMT2D, LPIN1, LPIN1, LPIN2, LPIN2, LPIN3, MAGED1, MAK, MAML1, MAML1, MAML1, MAML1, MAML2, MAML2, MAML3, MAML3, MAML3, MCIDAS,

MED1, MED1, MED1, MED1, MED1, MED1, MED6, MED 12, MED 12, MED 12, MED12L, MED 13, MED 14, MED 16, MED 17, MED 17, MED20, MED21, MED24, MED27, MED31, MEF2A, MMS19, MRTFA, MRTFB, MRTFB, MTA1, MTA1,

MTA1, MTA1, MTA2, MTA3, MTDH, MYCBP, MYOCD, MYOD1, MYSM1,

MYT1L, NACA, NCOA1, NCOA1, NCOA1, NCOA1, NCOA1, NCOA2, NCOA2, NCOA2, NCOA2, NCOA2, NCOA2, NCOA3, NCOA3, NCOA3, NCOA3, NCOA3, NCOA6, NCOA6, NCOA7, NEURODl, NEUROG3, NFE2L1, NKX2-2, NME2, NPAT, NPM1, NR1D1, NR1D2, NR1H2, NR1H3, NR1H3, NR1H4, NR1H5, NR1I2, NR1I3, NR3C1, NRBF2, NRIP1, NRIP1, NRIP1, NRL, NSD3, NUP98, NUPR1, PARK7, PCBD1, PDLIM1, PER2, PHF2, PKN1, PMF1, PMF1, PML, PML, PML, POU2AF1, POU3F1, POU3F2, POU3F2, POU4F1, POU4F2, POU5F1, PPARA, PPARD, PPARD, PPARG, PPARG, PPARG, PPARGC1A, PPARGC1A, PPARGC1A, PPARGC1A, PPARGC1 A, PPARGC1A, PPARGC1B, PPARGC1B, PPRC1, PRDM16, PRKCB, PRMT2, PRPF6, PRRX1, PSIP1, PSMC3IP, PSMC3IP, PSMD9, PSMD9, PUS1,

RAP2C, RARA, RARA, RARB, RARG, RBM14, RBM14, RBM39, RBPMS, RERE, REX04, RNF20, RRP1B, RUVBL1, RXRB, SCAND1, SERTAD2, SETD3, SFR1,

SFR1, SIX3, SLC30A9, SLC30A9, SMARCA2, SMARCA4, SMARCB1, SMARCB1, SMARCD3, SNW1, SNW1, SOX4, SOX11, SOX11, SOX12, SOX17, SP4, SRA1,

SRA1, SRA1, SRA1, SRA1, SRA1, SS18, SS18, SS18L1, SS18L2, SUB1, SUB1,

SUPT3, SUPT7L, TADA1, TADA1, T D A2A, TADA2B, TAD A3, TAD A3, TAD A3, TAFl, TAF5L, TAF6L, TAF6L, TAF7, TAF7, TAF7L, TAF9, TAFll, TAFll, TAF12, TCF3, TDRD3, TFAP2A, TFAP2A, TFAP2A, TFAP2B, TFAP2B, TFAP2B, TGFB1I1, THRA, THRAP3, THRAP3, THRAP3, THRB, TRIM24, TRIM24, TRIM28, TRIP4, TRIP4, TRRAP, TSG101, UBE2L3, UBE3A, USP16, USP21, USP22, USP22, UTF1, UTF1, VDR, VGLL2, WBP2, WBP2, WBP2NL, WDR77, WNT3A, WWC1, WWOX, WWTR1, WWTR1, YAF2, YAPl, YAPl, ZBTB18, ZCCHC12, ZCCHC12, ZCCHC18, ZMIZ2.

Applicable CRISPRa systems demonstrated to be capable of activating transcription in mammalian species include but are not limited to: VP64-p65-Rta (VPR), Synergistic Activation Mediator (SAM), Suntag, p300, and VP160.

One example of a transcriptional activator (or transactivator domain) is VP64. VP64 is made up of four copies of VP16, a viral protein sequence of 16 amino acids that is used for transcriptional activation. Embodiments of the invention include various forms of VP64, for example a nucleic acid comprising dCas9 and/or VP64, or plasmids or vectors that encode the dCas9 and/or VP64 genes. One non-limiting example includes pcDNA-dCas9-VP64 (Plasmid #47107, from Addgene). Additional elements can be present in the nucleic acid encoding dCas9 and/or VP64, as in for example lenti vector EFla-NLS-dCas9(N863)-VP64-2A-Blast-WPRE (Plasmid #61425 from Addgene), which additionally encodes a 2A Blast resistance marker. Another non-limiting example includes plasmid pLV hUbC-VP64 dCas9 VP64-T2A-GFP (Plasmid #59791 from Addgene) that co-expresses human optimized S. pyogenes dCas9 fused to two copies of VP64 and GFP.

Certain embodiments of the invention utilize the VP64-p65-Rta, or VPR, in which a VP64 transcriptional activator is joined to the C terminus of dCas9. In the dCas9-VPR protein, the transcription factors p65 and Rta are added to the C terminus of dCas9-VP64. Therefore, all three transcription factors are targeted to the same gene. The use of three transcription factors, as opposed to solely VP64, results in increased expression of targeted genes. dCas9-VPR can be used to increase expression of multiple genes within the same cell by putting multiple sgRNAs into the same cell.

In certain embodiments, the invention utilizes the Synergistic Activation Mediator (SAM) system. SAM makes use of not only VP64 but also sgRNA 2.0, which contains a sequence to recruit a viral protein fused to even more effectors (p65-hsfl). In one embodiment the SAM complex comprises dCas9-VP64, sgRNA, MS2-p65HSF-l. In one embodiment, the CRISPRa system comprises a nucleic acid encoding dCas9-VP64, a nucleic acid encoding MS2-p65-HSFl, and a genome-scale lentiviral SAM CRISPRa sgRNA library. Administering this type of CRISPRa system to a plurality of cells results in a highly diverse population of cells encompassing the entire sgRNA library.

The invention should be construed to work with any alternative activator, such as VP 16, VP160, p65AD, p300 or any other transcriptional activator.

The invention should also be construed to work with any dCas9/CRISPRa system or any other adaptor system known in the art, including but not limited to: 1) RNA Scaffolds, which also utilizes sgRNA 2.0 and recruits 3 viral proteins fused to VP64, 2) Suntag, which sports a protruding chain of 10 peptide epitopes that are recognized by an entourage of antibodies fused to VP64, 3) The epigenetic editor p300, which deposits activating H3K27ac. 4) VP 160, which is also known as CRISPR-on and has ten times the VPs of VP 16, and 5) VP64-dCas9-BFP-VP64, which makes use of that much neglected N-terminus.

The CRISPRa systems disclosed herein can be used with an sgRNA library. With regard to any and all sgRNA libraries disclosed herein, it should be understood by one of ordinary skill in the art that when it is stated that the library comprises at least one sgRNA, it should be construed that the library can comprise one or more sgRNAs, all sgRNAs in the library, and and all integer values and numerical ranges of sgRNAs there between. For example, an sgRNA library comprising a total of 219 sgRNAs (e.g. SEQ ID NOs. 193-411) can comprise one sgRNA, all 219 sgRNAs, or and any and all integer values between 1 and 219. In other words, the library could include 1, 10, 20, 50, 100, 150, 200, or 219 sgRNAs and any and all values in between.

CRISPRa screen assisted rational design of off-the-shelf viral immune gene therapies Provided herein is a method of developing an off-the-shelf viral immune gene therapy. The method utilizes an in vivo CRISPRa screen to unbiasedly pinpoint the genetic factors that enhance host immune surveillance and tumor destruction (“rational design”, or “immune gene therapy rationalization”). In one embodiment, the method comprises administering an APCM sgRNA library (e.g. SEQ ID NOs: 86-192) to a cell (e.g. a cancer cell) or population of cells. “APCM" refers to an approach of multiplexed immune gene therapy (abbreviated for Antigen Presentation CRISPR Manipulation; Antigen Presentation and Co-stimulation Manipulation; and/or Antigen Presentation, Co stimulation and Migration) wherein a plurality of APCM genes are activited by the CRISPRa system using sgRNAs specific for these antigen presentation genes, or by overexpression with cDNA expressing ORFs, or by other gene activation / expression / delivery approaches, or by the combination(s) of above. The cel!(s) are cultured in vitro , thereby generating a modified cell or population of cells, then injected into a mammal (e.g. a mouse). Tumor cells are collected from the mammal and analyzed to determine which genes are depleted (e.g. sgRNA depletion and enrichment readout by Illumina sequencing). Specifically, sgRNAs that that are depleted by the mammal’s immune system in the resulting tumors, identifying the genes that confer immune recognition of tumor cells. This novel screening approach is based on immune rejection, e.g. identifies the strongest immune factors. By using this method, one can identify which combination of genes should be targeted, e.g. those that are rejected by the immune system, rather than guessing. Genes that are depleted by the method are highly immunogenic and work the best. This method can be used for different tumor/cancer types, as each tumor/cancer type may involve a different set of genes that are immunogenic. Thus, different therapies can be designed to target different combinations of genes from different types of cancers.

In one aspect, the invention provides a method of developing a cancer therapy (e.g. a rationalized immune gene therapy). The method comprises administering a CRISPR activation (CRISPRa) system comprising a sgRNA library to a cancer cell, thereby generating a modified cell, administering the modified cell to a mammal whereby the mammal develops cancer, determining the genes that are depleted in the cancer cell from the mammal, and designing a cancer therapy that targets the depleted genes (e.g immune promoting genes).

In certain embodiments, the sgRNA library comprises the nucleotide sequences set forth in SEQ ID NOs. 86-192 (Fig. 7). In certain embodiments, the sgRNA library comprises an AAV-APCM library (AAV-Apcm) designed/optimized specifically for human genes. Such gRNAs were designed to target the same set of immune genes as above, yielding a library of 219 sgRNAs (SEQ ID NOs: 193-411) (Figs. 8A-8B).

In certain embodiments, the mammal is a mouse. Other mammals can be used include but are not limited to dogs, cats, pigs, and humans.

In certain embodiments, the CRISPRa system comprises a vector comprising the nucleotide sequence set forth in SEQ ID NO: 1.

Determining the genes that are depleted in the cancer cell can be achieved by any means known to one of ordingary skill in the art, for example, nucleotide sequencing and analysis.

In certain embodiments, designing a cancer therapy that targets the depleted genes comprises packaging the open reading frames (ORFs) of the depleted genes in a vector. In certain embodiments, the vector is an adeno-associated viral (AAV) vector.

Compositions

Certain aspects of the invention comprise vectors and compositions comprising vectors. In certain embodiments, the vectors and compositons are useful for treating cancer.

In one aspect, the invention provides an off-the-shelf therapy useful for treating cancer ( e.g . triple negative breast cancer). In one embodiment, the therapy comprises CLC4, which targets CD80, Light, CXCL10, and 4-1BBL genes. In one embodiment, the therapy comprises CLC4I, which targets CD80, Light, CXCL10, 4-1BBL, and IFNG. In one embodiment, the therapy comprises CLC4G, which targets CD80, Light, CXCL10, 4- 1BBL, and GITRL. These therapies can be in the form of MAEGI (CRISPRa-based pooled activation), direct ORF -based viral immune gene therapy (AAV expressing concatenated ORFs), or other forms.

In one aspect, the invention provides a composition comprising a vector comprising an open reading frame (ORF) of at least one gene selected from the group consisting of CD80, Light, CXCL10, 4-1BBL, GITRL, IL2, IL-23, and IFNg. The invention should be construed to include ORFs from any number or any combination of the genes selected from the group consisting of CD80, Light, CXCL10, 4-1BBL, GITRL, IL2, IL-23, and IFNg. For example, the vector can comprise one or more ORFs from one or more of the aforementioned genes. The vector can comprise one or more ORFs from any combination of the gene selected from the group consisting of CD80, Light,

CXCL10, 4-1BBL, GITRL, IL2, IL-23, and IFNg. For example, the vector can comprise ORFs from one, two, three, four, five, six, seven, or eight genes selected from the group consisting of CD80, Light, CXCL10, 4-1BBL, GITRL, IL2, IL-23, and IFNg.

In one aspect, the invention provides a composition comprising a vector comprising the open reading frames (ORFs) of CD80, Light, CXCL10, 4-1BBL and IFNG genes. In another aspect, the invention provides a vector comprising the open reading frames (ORFs) of CD80, Light, CXCL10, and 4-1BBL. In certain embodiments, vector is an AAV vector. In certain embodiments, the vector comprises the nucleotide sequence set forth in SEQ ID NO: 2 or SEQ ID NO: 3.

In one aspect, the invention provides a composition comprising a vector comprising the open reading frames (ORFs) of IL-23, 4-1BBL, and IFNg genes. In one aspect, the invention provides a composition comprising a vector comprising the open reading frames (ORFs) of IL-23, Light, CXCL10, 4-1BBL and IFNg genes. In one aspect, the invention provides a composition comprising a vector comprising the open reading frames (ORFs) of IFNg, CD80, Light, and 4-1BBL and genes. In one aspect, the invention provides a composition comprising a vector comprising the open reading frames (ORFs) of IL-23, 4-1BBL, and IFNg genes. In one aspect, the invention provides a composition comprising a vector comprising the open reading frames (ORFs) of CD80, 41BBL, IL23 and IFNg genes.

In one aspect, the invention provides a composition comprising an adenoviral- based dSpCas9 CRISPRa system that utilizes a single vector to deliver all CRISPRa components. In one embodiment, the vector comprises pGW029 (SEQ ID NO: 4). In one embodiment, the vector comprises pGW035 (SEQ ID NO: 5), In one embodiment, the vector comprises pGW063 (SEQ ID NO: 6),

In another aspect, the invention provides a composition comprising a dSaCas9- based AIO (all-in-one) CRISPRa system, which uses a single AAV vector to deliver all CRISPRa components. In one embodiment, the vector comprises pZB3 (SEQ ID NO:

10).

In another aspect, the invention provides a composition comprising an AAV based two-vector system with dSaCas9. In this system, the dSaCas9 enzyme is on a separate vector from the other CRISPRa components. In one embodiment, the vector which contains the dSaCas9 is pGW060 (SEQ ID NO: 12) and the vector that contains the other CRISPRa components is pGW047 (SEQ ID NO: 11).

In another aspect, the invention provides a composition comprising an AAV based two-vector system with dSpCas9. In this system, the dSpCas9 enzyme is on a separate vector from the other CRISPRa components. In one embodiment, the vector which contains the dSpCas9 is pGWOl lb (SEQ ID NO: 56) and the vector that contains the other CRISPRa components is pGW045 (SEQ ID NO: 13).

In another aspect, the invention provides a composition comprising an AAV based two-vector system with dCasl2/dCpfl. In certain embodiments, the composition comprises one or more vectors selected from the group consisting of pRCl 19 (SEQ ID NO: 14), pRC120 (SEQ ID NO: 15), pRC121b(SEQ ID NO: 16), pRC124(SEQ ID NO: 17), and pRC126 (SEQ ID NO: 18).

In one aspect, the invention includes a vector comprising SEQ ID NO: 1. In another aspect, the invention includes a vector comprising SEQ ID NO: 2. In another aspect, the invention includes a vector comprising SEQ ID NO: 3. In another aspect, the invention includes a vector comprising SEQ ID NO: 4. In another aspect, the invention includes a vector comprising SEQ ID NO: 5. In another aspect, the invention includes a vector comprising SEQ ID NO: 6. In another aspect, the invention includes a vector comprising SEQ ID NO: 7. In another aspect, the invention includes a vector comprising SEQ ID NO: 8. In another aspect, the invention includes a vector comprising SEQ ID NO: 9. In another aspect, the invention includes a vector comprising SEQ ID NO: 10. In another aspect, the invention includes a vector comprising SEQ ID NO: 11. In another aspect, the invention includes a vector comprising SEQ ID NO: 12. In another aspect, the invention includes a vector comprising SEQ ID NO: 13. In another aspect, the invention includes a vector comprising SEQ ID NO: 14. In another aspect, the invention includes a vector comprising SEQ ID NO: 15. In another aspect, the invention includes a vector comprising SEQ ID NO: 16. In another aspect, the invention includes a vector comprising SEQ ID NO: 17. In another aspect, the invention includes a vector comprising SEQ ID NO: 18. In another aspect, the invention includes a vector comprising SEQ ID NO: 24. In another aspect, the invention includes a vector comprising SEQ ID NO: 25. In another aspect, the invention includes a vector comprising SEQ ID NO: 26. In another aspect, the invention includes a vector comprising SEQ ID NO: 27. In another aspect, the invention includes a vector comprising SEQ ID NO: 28. In another aspect, the invention includes a vector comprising SEQ ID NO: 29. In another aspect, the invention includes a vector comprising SEQ ID NO: 30. In another aspect, the invention includes a vector comprising SEQ ID NO: 31. In another aspect, the invention includes a vector comprising SEQ ID NO: 32. In another aspect, the invention includes a vector comprising SEQ ID NO: 33. In another aspect, the invention includes a vector comprising SEQ ID NO: 34. In another aspect, the invention includes a vector comprising SEQ ID NO: 35. In another aspect, the invention includes a vector comprising SEQ ID NO: 36. In another aspect, the invention includes a vector comprising SEQ ID NO: 37. In another aspect, the invention includes a vector comprising SEQ ID NO: 38. In another aspect, the invention includes a vector comprising SEQ ID NO: 39. In another aspect, the invention includes a vector comprising SEQ ID NO: 40. In another aspect, the invention includes a vector comprising SEQ ID NO: 41. In another aspect, the invention includes a vector comprising SEQ ID NO: 42. In another aspect, the invention includes a vector comprising SEQ ID NO: 43. In another aspect, the invention includes a vector comprising SEQ ID NO: 44. In another aspect, the invention includes a vector comprising SEQ ID NO: 45. In another aspect, the invention includes a vector comprising SEQ ID NO: 46. In another aspect, the invention includes a vector comprising SEQ ID NO: 47. In another aspect, the invention includes a vector comprising SEQ ID NO: 48. In another aspect, the invention includes a vector comprising SEQ ID NO: 49. In another aspect, the invention includes a vector comprising SEQ ID NO: 50. In another aspect, the invention includes a vector comprising SEQ ID NO: 51. In another aspect, the invention includes a vector comprising SEQ ID NO: 52. In another aspect, the invention includes a vector comprising SEQ ID NO: 53. In another aspect, the invention includes a vector comprising SEQ ID NO: 54. In another aspect, the invention includes a vector comprising SEQ ID NO: 55. In another aspect, the invention includes a vector comprising SEQ ID NO: 56.

In certain aspects, the invention includes an AAV-ORF immune gene therapy construct. In certain aspects, the invention includes any of the contstructs/ vectors depicted in Fig. 15. In certain aspects, the invention includes a vector comprising a nucleotide sequence encoding at least one of the following components: Ori, flOri,

AmpR promoter, AmpR, AAV-ITR, EFS core promoter, LTR promoter, EFS-LTR promoter, T2A, P2A, E2A, WPRE, short-Poly A, ORF-Cd80, ORF-Light, ORF-CxcllO, ORF-41BBL, ORF-IFNg, ORF-I12, ORF-Gitrl, ORF-I123, ORF-hIFNg, ORF-hIL23, ORF-h41BBL, ORF-hLIGHT(remove-EQLI), CGKRK. In certain aspects, the invention includes a vector comprising the nucleotide sequence set forth in any one of SEQ ID NOs: 59-85.

In certain aspects, the invention includes a vector comprising a nucleotide sequence encoding at least one ORF selected from the group consisting of ORF-Light, ORF-CxcllO, ORF-41BBL, ORF-IFNg, ORF-I12, ORF-Gitrl, ORF-I123, ORF-hIFNg, ORF-hIL23, ORF-h41BBL, ORF-hLIGHT, or any combination thereof. In certain embodiments, the vector comprises one, two, three, four, five, six, seven, eight, nine, ten, or eleven ORF sequences selected from the group consisting of ORF-Light, ORF-CxcllO, ORF -41 BBL, ORF -IFNg, ORF-I12, ORF-Gitrl, ORF-I123, ORF-hIFNg, ORF-ML23, ORF-h41BBL, and ORF-hLIGHT. In certain embodiments, the vector comprises at least one nucleotide sequence selected from the group consisting of SEQ ID NOs: 73-84. In certain embodiments, the vector comprises one, two, three, four, five, six, seven, eight, nine, ten, or eleven nucleotide sequences selected from the group consisting of SEQ ID NOs: 73-84.

Tolerable variations of any one of the vectors or component parts will be known to those of skill in the art. For example, in some embodiments the vector or component part comprises a nucleic acid sequence that has at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to any one of the nucleic acid sequence set forth in SEQ ID NO: 1-18 or 24-85.

Methods of Treatment

The present invention includes methods for treating or preventing cancer in a subject comprising administering to the subject a therapeutically effective amount of any of the compositions disclosed herein. This composition can be utilized as a prophylactic treatment, a therapeutic treatment, a personalized, subject-specific treatment, and/or a method of turning a ‘cold’ tumor into a ‘hot’ tumor, thus making it more susceptible to immunotherapy.

One aspect of the invention includes a method of treating cancer in a subject in need thereof comprising administering to the subject a therapeutically effective amount of a composition comprising a vector comprising a CRISPRa system, wherein the CRISPRa system increases expression of at least one endogenous gene, thus treating the cancer in the subject.

The vector can be any vector that can carry the gene, including but not limited to, standard viral vectors, chimeric viral vectors, other viral vectors, bacterial vectors, yeast vectors, DNA vectors, mRNA, protein carriers, nanomaterials, or other delivery vehicles. Applicable standard viral vectors for delivery include the vectors from the following types of viruses: dsDNA viruses ( e.g . Adenoviruses, Herpesviruses, Poxviruses), ssDNA viruses (+ strand or "sense") DNA (e.g. Parvoviruses), dsRNA viruses ( e.g . Reoviruses), (+)ssRNA viruses (+ strand or sense) RNA (e.g. Picomaviruses, Togaviruses), (-)ssRNA viruses (- strand or antisense) RNA (e.g. Orthomyxoviruses, Rhabdoviruses), ssRNA-RT viruses (+ strand or sense) RNA with DNA intermediate in life-cycle (e.g. Retroviruses), and dsDNA-RT viruses DNA with RNA intermediate in life-cycle (e.g. Hepadnaviruses).

In certain embodiments of the invention, the cells are packaged into an AAV vector. Applicable AAV serotypes include, but are not limited to, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV 12, artificial variants such as AAV.rhlO, AAV.rh32/33, AAV.rh43, AAV.rh64Rl, rAAV2-retro, AAV- DJ, AAV-PHP.B, AAV-PHP.S, AAV-PHP.eB, or other engineered versions of AAV. In one embodiment, the AAV vector is AAV9. In one embodiment, the CRISPRa system is cloned into an AAV vector.

The cell or cells utilized in the invention can be from any source known to one of ordinary skill in the art. The cells of the invention may be autologous, allogeneic or xenogeneic with respect to the subject undergoing treatment. In some embodiments, the cell is from a cancer cell line. In some embodiments, the cell is from the subject. In some embodiments, the cell from the subject is a cancer cell. In further embodiments, the cancer cell is from a tumor. In some embodiments, the subject is a mammal. In further embodiments, the subject is a human.

The cells or vectors of the present invention may be administered in a manner appropriate to the disease to be treated or prevented. The quantity and frequency of administration will be determined by such factors as the condition of the patient, and the type and severity of the patient’s disease, although appropriate dosages may be determined by clinical trials. Cells or vectors of the invention can be administered in dosages and routes and at times to be determined in appropriate pre-clinical and clinical experimentation and trials. Cell and vector compositions may be administered multiple times at various dosages. Administration of the cells or vectors of the invention may be combined with other methods useful to treat the desired disease or condition as determined by those of skill in the art. In one embodiment, administering the therapeutically effective amount of the composition comprises a one dose, a two dose, a three dose, a four dose, or a multi-dose treatment. The administration of the modified cells or vectors of the invention may be carried out in any convenient manner known to those of skill in the art. In one embodiment, the cells are administered intratum orally. The invention includes compositions and methods for treating cancer. Types of cancer that can be treated include, but are not limited to, Acute Lymphoblastic Leukemia (ALL), Acute Myeloid Leukemia (AML), Adrenocortical Carcinoma, AIDS-Related Cancers, Kaposi Sarcoma, AIDS-Related Lymphoma, Primary CNS Lymphoma, Anal Cancer, Appendix Cancer (Gastrointestinal Carcinoid Tumors), Astrocytomas, Atypical Teratoid/Rhabdoid Tumor, Brain Cancer, Basal Cell Carcinoma of the Skin, Bile Duct Cancer, Bladder Cancer, Bone Cancer (includes Ewing Sarcoma and Osteosarcoma and Malignant Fibrous Histiocytoma), Brain Tumors, Breast Cancer, Bronchial Tumors, Burkitt Lymphoma, Non-Hodgkin Lymphoma, Carcinoid Tumors, Carcinoma of Unknown Primary, Cardiac (Heart) Tumors, Embryonal Tumors, Germ Cell Tumor, Primary CNS Lymphoma, Cervical Cancer, Cholangiocarcinoma, Chordoma, Chronic Lymphocytic Leukemia (CLL), Chronic Myelogenous Leukemia (CML), Chronic Myeloproliferative Neoplasms, Colorectal Cancer, Craniopharyngioma, Cutaneous T-Cell Lymphoma (Mycosis Fungoides and Sezary Syndrome), Ductal Carcinoma In Situ (DCIS), Endometrial Cancer, Ependymoma, Esophageal Cancer, Esthesioneuroblastoma, Ewing Sarcoma, Extracranial Germ Cell Tumor, Eye Cancer, Intraocular Melanoma, Fallopian Tube Cancer, Fibrous Histiocytoma of Bone, Osteosarcoma, Gallbladder Cancer, Gastric Cancer, Stomach Cancer, Gastrointestinal Carcinoid Tumor, Gastrointestinal Stromal Tumors (GIST), Central Nervous System Germ Cell Tumors, Extracranial Germ Cell Tumors, Extragonadal Germ Cell Tumors, Ovarian Germ Cell Tumors, Testicular Cancer, Gestational Trophoblastic Disease, Hairy Cell Leukemia, Head and Neck Cancer, Heart Tumors, Hepatocellular (Liver) Cancer, Histiocytosis (Langerhans Cell), Hodgkin Lymphoma, Hypopharyngeal Cancer, Intraocular Melanoma, Islet Cell Tumors, Pancreatic Neuroendocrine Tumors, Kidney Cancer, Renal Cell Cancer, Langerhans Cell Histiocytosis, Laryngeal Cancer, Leukemia, Lip and Oral Cavity Cancer, Liver Cancer, Lung Cancer (Non-Small Cell and Small Cell), Lymphoma, Male Breast Cancer, Malignant Fibrous Histiocytoma of Bone and Osteosarcoma, Melanoma, Intraocular (Eye) Melanoma, Merkel Cell Carcinoma (Skin Cancer), Malignant Mesothelioma, Metastatic Cancer, Metastatic Squamous Neck Cancer with Occult Primary, Midline Tract Carcinoma With NUT Gene Changes, Mouth Cancer, Multiple Endocrine Neoplasia Syndromes, Multiple Myeloma/Plasma Cell Neoplasms, Mycosis Fungoides (Lymphoma), Myelodysplastic Syndromes, Myelodysplastic/ Myeloproliferative Neoplasms, Nasal Cavity and Paranasal Sinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Non-Small Cell Lung Cancer, Oral Cancer, and Oropharyngeal Cancer, Ovarian Cancer, Pancreatic Cancer, Papillomatosis, Paraganglioma, Paranasal Sinus and Nasal Cavity Cancer, Parathyroid Cancer, Penile Cancer, Pharyngeal Cancer, Pheochromocytoma, Pituitary Tumor, Plasma Cell Neoplasm/Multiple Myeloma, Pleuropulmonary Blastoma, Primary Central Nervous System (CNS) Lymphoma, Primary Peritoneal Cancer, Prostate Cancer, Rectal Cancer Recurrent Cancer, Retinoblastoma, Rhabdomyosarcoma, Salivary Gland Cancer,

Sarcoma, Vascular Tumors, Uterine Sarcoma, Sezary Syndrome (Lymphoma), Small Cell Lung Cancer, Small Intestine Cancer, Soft Tissue Sarcoma, Squamous Cell Carcinoma, Stomach (Gastric) Cancer, Throat Cancer, Thymoma, Thymic Carcinoma, Thyroid Cancer, Transitional Cell Cancer of the Renal Pelvis and Ureter, Carcinoma of Unknown Primary, Ureter and Renal Pelvis, Transitional Cell Cancer, Urethral Cancer, Uterine Cancer, Vaginal Cancer, Vulvar Cancer, Wilms Tumor, and combinations thereof.

Certain embodiments of the invention further comprise administering an additional treatment to the subject. Certain embodiments of the invention include treating the subject with a combination of a composition of the present invention and an additional treatment. Examples of additional treatments include but are not limited to, chemotherapy, radiation, surgery, medication, immune checkpoint inhibitors, immune checkpoint blockade (ICB) antibodies, immune checkpoint inhibitors that block CTLA-4 orPDl, anti-CTLA4 monoclonal antibody, anti -PD 1 monoclonal antibody, anti-PD-Ll monoclonal antibody, adoptive cell transfer, human recombinant cytokines, cancer vaccines, immunotherapy, targeted therapy, hormone therapy, stem cell transplant, precision medicine, non-specific immunotherapy ( e.g . cytokines and chemokines, such as IL-2, IL-7, IL-12, IL-15, IL-18, IL-23, IFNa, IFNb, IFNg, TNFa), oncolytic virus therapy, cell therapy (e.g. adoptive transfer of TILs, TCR-T, CAR-T, CAR-NK, CAR- macrophage, or other forms of naive, patient-isolated or engineered primary cells), cancer vaccines (e.g. conventional DC vaccine), Ipilimumab (Yervoy), Nivolumab (Opdivo), Pembrolizumab (Keytruda), Atezolizumab (Tecentriq), Avelumab (Bavencio), Durvalumab (Imfinzi), Anti-LAG-3, anti-TIMl, Anti-TIM3, Anti-CSF-R, IDO inhibitor, OX-40 agonist, GITR agonist, CD80 agonist, CD86 agonist, ICOS agonist, ICOSLG agonist, CD276 agonist, VTCN1 agonist, TNFSF14 agonist, TNFSF9 agonist, TNFSF4 agonist, CD70 agonist, CD40 agonist, LGALS9 agonist, CD80 inhibitor, CD86 inhibitor, ICOS inhibitor, ICOSLG inhibitor, CD276 inhibitor, VTCN1 inhibitor, TNFSF14 inhibitor, TNFSF9 inhibitor, TNFSF4 inhibitor, CD70 inhibitor, CD40 inhibitor,

LGALS9 inhibitor, TLR9 agonist, CD20 antibody, CD80 antibody, TIGIT antibody, B7- HI antibody, B7-H2 antibody, B7-H3 antibody, B7-H4 antibody, CD28 antibody, CD47 antibody, anti-BTLA, anti-Galetin9, anti-IL15R, anti-GD2. In some embodiments the monoclonal antibody is fully human, humanized or chimeric.

In certain embodiments, administering a composition of the present invention alters the tumor microenvironment. In certain embodiments, administering the composition augments host immune responses against established tumors.

Introduction of Nucleic Acids

In certain embodiments an expression system is used for the introduction of gRNAs and (d)Cas9 proteins into the cells of interest. Typically employed options include but are not limited to plasmids and viral vectors such as adeno-associated virus (AAV) vector, adenoviral vector or lentivirus vector.

Methods of introducing nucleic acids into a cell include physical, biological and chemical methods. Physical methods for introducing a polynucleotide, such as RNA, into a host cell include calcium phosphate precipitation, lipofection, particle bombardment, microinjection, electroporation, and the like. RNA can be introduced into target cells using commercially available methods which include electroporation (Amaxa Nucleofector-II (Amaxa Biosystems, Cologne, Germany)), (ECM 830 (BTX) (Harvard Instruments, Boston, Mass.) or the Gene Pulser II (BioRad, Denver, Colo.), Multiporator (Eppendort, Hamburg Germany). RNA can also be introduced into cells using cationic liposome mediated transfection using lipofection, using polymer encapsulation, using peptide mediated transfection, or using biolistic particle delivery systems such as “gene guns” (see, for example, Nishikawa, et al. Hum Gene Then, 12(8):861-70 (2001).

Biological methods for introducing a polynucleotide of interest into a host cell include the use of DNA and RNA vectors. Viral vectors have become the most widely used method for introducing genes into mammalian, e.g., human cells. Other viral vectors can include as listed above. See, for example, U.S. Pat. Nos. 5,350,674 and 5,585,362.

Chemical means for introducing a polynucleotide into a host cell include colloidal dispersion systems, such as macromolecule complexes, nanocapsules, microspheres, beads, and lipid-based systems including oil-in-water emulsions, micelles, mixed micelles, and liposomes. An exemplary colloidal system for use as a delivery vehicle in vitro and in vivo is a liposome (e.g., an artificial membrane vesicle).

Lipids suitable for use can be obtained from commercial sources. For example, dimyristyl phosphatidylcholine (“DMPC”) can be obtained from Sigma, St. Louis, MO; dicetyl phosphate (“DCP”) can be obtained from K & K Laboratories (Plainview, NY); cholesterol (“Choi”) can be obtained from Calbiochem-Behring; dimyristyl phosphatidylglycerol (“DMPG”) and other lipids may be obtained from Avanti Polar Lipids, Inc. (Birmingham, AL). Stock solutions of lipids in chloroform or chloroform/methanol can be stored at about -20°C. Chloroform is used as the only solvent since it is more readily evaporated than methanol. “Liposome” is a generic term encompassing a variety of single and multilamellar lipid vehicles formed by the generation of enclosed lipid bilayers or aggregates. Liposomes can be characterized as having vesicular structures with a phospholipid bilayer membrane and an inner aqueous medium. Multilamellar liposomes have multiple lipid layers separated by aqueous medium. They form spontaneously when phospholipids are suspended in an excess of aqueous solution. The lipid components undergo self-rearrangement before the formation of closed structures and entrap water and dissolved solutes between the lipid bilayers (Ghosh et al., 1991 Glycobiology 5: 505-10). However, compositions that have different structures in solution than the normal vesicular structure are also encompassed. For example, the lipids may assume a micellar structure or merely exist as nonuniform aggregates of lipid molecules. Also contemplated are lipofectamine-nucleic acid complexes.

Regardless of the method used to introduce exogenous nucleic acids into a host cell or otherwise expose a cell to the inhibitor of the present invention, in order to confirm the presence of the nucleic acids in the host cell, a variety of assays may be performed. Such assays include, for example, “molecular biological” assays well known to those of skill in the art, such as Southern and Northern blotting, RT-PCR and PCR; “biochemical” assays, such as detecting the presence or absence of a particular peptide, e.g., by immunological means (ELIS As and Western blots) or by assays described herein to identify agents falling within the scope of the invention.

Moreover, the nucleic acids may be introduced by any means, such as transducing the cells, transfecting the cells, and electroporating the cells. One nucleic acid may be introduced by one method and another nucleic acid may be introduced into the cell by a different method.

RNA

In one embodiment, the nucleic acids introduced into the cell are RNA. In another embodiment, the RNA is mRNA that comprises in vitro transcribed RNA or synthetic RNA. The RNA is produced by in vitro transcription using a polymerase chain reaction (PCR)-generated template. DNA of interest from any source can be directly converted by PCR into a template for in vitro mRNA synthesis using appropriate primers and RNA polymerase. The source of the DNA can be, for example, genomic DNA, plasmid DNA, phage DNA, cDNA, synthetic DNA sequence or any other appropriate source of DNA.

PCR can be used to generate a template for in vitro transcription of mRNA which is then introduced into cells. Methods for performing PCR are well known in the art. Primers for use in PCR are designed to have regions that are substantially complementary to regions of the DNA to be used as a template for the PCR. “Substantially complementary”, as used herein, refers to sequences of nucleotides where a majority or all of the bases in the primer sequence are complementary, or one or more bases are non complementary, or mismatched. Substantially complementary sequences are able to anneal or hybridize with the intended DNA target under annealing conditions used for PCR. The primers can be designed to be substantially complementary to any portion of the DNA template. For example, the primers can be designed to amplify the portion of a gene that is normally transcribed in cells (the open reading frame), including 5' and 3' UTRs. The primers can also be designed to amplify a portion of a gene that encodes a particular domain of interest. In one embodiment, the primers are designed to amplify the coding region of a human cDNA, including all or portions of the 5' and 3' UTRs. Primers useful for PCR are generated by synthetic methods that are well known in the art. “Forward primers” are primers that contain a region of nucleotides that are substantially complementary to nucleotides on the DNA template that are upstream of the DNA sequence that is to be amplified. “Upstream” is used herein to refer to a location 5, to the DNA sequence to be amplified relative to the coding strand. “Reverse primers” are primers that contain a region of nucleotides that are substantially complementary to a double-stranded DNA template that are downstream of the DNA sequence that is to be amplified. “Downstream” is used herein to refer to a location 3' to the DNA sequence to be amplified relative to the coding strand.

Chemical structures that have the ability to promote stability and/or translation efficiency of the RNA may also be used. The RNA preferably has 5' and 3' UTRs. In one embodiment, the 5' UTR is between zero and 3000 nucleotides in length. The length of 5' and 3' UTR sequences to be added to the coding region can be altered by different methods, including, but not limited to, designing primers for PCR that anneal to different regions of the UTRs. Using this approach, one of ordinary skill in the art can modify the 5' and 3' UTR lengths required to achieve optimal translation efficiency following transfection of the transcribed RNA.

The 5' and 3' UTRs can be the naturally occurring, endogenous 5' and 3' UTRs for the gene of interest. Alternatively, UTR sequences that are not endogenous to the gene of interest can be added by incorporating the UTR sequences into the forward and reverse primers or by any other modifications of the template. The use of UTR sequences that are not endogenous to the gene of interest can be useful for modifying the stability and/or translation efficiency of the RNA. For example, it is known that AU-rich elements in 3' UTR sequences can decrease the stability of mRNA. Therefore, 3' UTRs can be selected or designed to increase the stability of the transcribed RNA based on properties of UTRs that are well known in the art.

In one embodiment, the 5' UTR can contain the Kozak sequence of the endogenous gene. Alternatively, when a 5' UTR that is not endogenous to the gene of interest is being added by PCR as described above, a consensus Kozak sequence can be redesigned by adding the 5' UTR sequence. Kozak sequences can increase the efficiency of translation of some RNA transcripts, but does not appear to be required for all RNAs to enable efficient translation. The requirement for Kozak sequences for many mRNAs is known in the art. In other embodiments the 5' UTR can be derived from an RNA virus whose RNA genome is stable in cells. In other embodiments various nucleotide analogues can be used in the 3' or 5' UTR to impede exonuclease degradation of the mRNA.

To enable synthesis of RNA from a DNA template, a promoter of transcription should be attached to the DNA template upstream of the sequence to be transcribed.

When a sequence that functions as a promoter for an RNA polymerase is added to the 5' end of the forward primer, the RNA polymerase promoter becomes incorporated into the PCR product upstream of the open reading frame that is to be transcribed. In one embodiment, the promoter is a T7 polymerase promoter, as described elsewhere herein. Other useful promoters include, but are not limited to, T3 and SP6 RNA polymerase promoters. Consensus nucleotide sequences for T7, T3 and SP6 promoters are known in the art.

In one embodiment, the mRNA has a cap on the 5' end and a 3' poly(A) tail which determine ribosome binding, initiation of translation and stability mRNA in the cell. On a circular DNA template, for instance, plasmid DNA, RNA polymerase produces a long concatameric product which may not be suitable for expression in eukaryotic cells. The transcription of plasmid DNA linearized at the end of the 3' UTR results in normal sized mRNA which may not be effective in eukaryotic transfection even if it is polyadenylated after transcription.

On a linear DNA template, phage T7 RNA polymerase can extend the 3' end of the transcript beyond the last base of the template (Schenborn and Mierendorf, Nuc Acids Res., 13:6223-36 (1985); Nacheva and Berzal-Herranz, Eur. J. Biochem., 270:1485-65 (2003).

The conventional method of integration of polyA/T stretches into a DNA template is by molecular cloning. However polyA/T sequence integrated into plasmid DNA can cause plasmid instability, which is why plasmid DNA templates obtained from bacterial cells are often highly contaminated with deletions and other aberrations. This makes cloning procedures not only laborious and time consuming but often not reliable. That is why a method which allows construction of DNA templates with polyA/T 3' stretch without cloning highly desirable.

The polyA/T segment of the transcriptional DNA template can be produced during PCR by using a reverse primer containing a polyT tail, such as 100T tail (size can be 50-5000 T), or after PCR by any other method, including, but not limited to, DNA ligation or in vitro recombination. Poly(A) tails also provide stability to RNAs and reduce their degradation. Generally, the length of a poly(A) tail positively correlates with the stability of the transcribed RNA. In one embodiment, the poly(A) tail is between 100 and 5000 adenosines.

Poly(A) tails of RNAs can be further extended following in vitro transcription with the use of a poly (A) polymerase, such as E. coli poly A polymerase (E-PAP). In one embodiment, increasing the length of a poly(A) tail from 100 nucleotides to between 300 and 400 nucleotides results in about a two-fold increase in the translation efficiency of the RNA. Additionally, the attachment of different chemical groups to the 3' end can increase mRNA stability. Such attachment can contain modified/artificial nucleotides, aptamers and other compounds. For example, ATP analogs can be incorporated into the poly(A) tail using poly(A) polymerase. ATP analogs can further increase the stability of the RNA.

5' caps also provide stability to RNA molecules. In a preferred embodiment,

RNAs produced by the methods disclosed herein include a 5' cap. The 5' cap is provided using techniques known in the art and described herein (Cougot, et ah, Trends in Biochem. Sci., 29:436-444 (2001); Stepinski, et ah, RNA, 7:1468-95 (2001); Elango, et ah, Biochim. Biophys. Res. Commun., 330:958-966 (2005)). The RNAs produced by the methods disclosed herein can also contain an internal ribosome entry site (IRES) sequence. The IRES sequence may be any viral, chromosomal or artificially designed sequence which initiates cap-independent ribosome binding to mRNA and facilitates the initiation of translation. Any solutes suitable for cell electroporation, which can contain factors facilitating cellular permeability and viability such as sugars, peptides, lipids, proteins, antioxidants, and surfactants can be included.

In some embodiments, the RNA is electroporated into the cells, such as in vitro transcribed RNA.

The methods also provide the ability to control the level of expression over a wide range by changing, for example, the promoter or the amount of input RNA, making it possible to individually regulate the expression level. Furthermore, the PCR-based technique of mRNA production greatly facilitates the design of the mRNAs with different structures and combination of their domains.

One advantage of RNA transfection methods of the invention is that RNA transfection is essentially transient and vector-free. A RNA transgene can be delivered to a lymphocyte and expressed therein following a brief in vitro cell activation, as a minimal expressing cassette without the need for any additional viral sequences. Under these conditions, integration of the transgene into the host cell genome is unlikely. Cloning of cells is not necessary because of the efficiency of transfection of the RNA and its ability to uniformly modify the entire lymphocyte population.

Genetic modification of cells with in vvV/o-tran scribed RNA (IVT-RNA) makes use of two different strategies both of which have been successively tested in various animal models. Cells are transfected with in vitro- transcribed RNA by means of lipofection or electroporation. It is desirable to stabilize IVT-RNA using various modifications in order to achieve prolonged expression of transferred IVT-RNA.

Some IVT vectors are known in the literature which are utilized in a standardized manner as template for in vitro transcription and which have been genetically modified in such a way that stabilized RNA transcripts are produced. Currently protocols used in the art are based on a plasmid vector with the following structure: a 5' RNA polymerase promoter enabling RNA transcription, followed by a gene of interest which is flanked either 3' and/or 5' by untranslated regions (UTR), and a 3' polyadenyl cassette containing 50-70 A nucleotides. Prior to in vitro transcription, the circular plasmid is linearized downstream of the polyadenyl cassette by type II restriction enzymes (recognition sequence corresponds to cleavage site). The polyadenyl cassette thus corresponds to the later poly(A) sequence in the transcript. As a result of this procedure, some nucleotides remain as part of the enzyme cleavage site after linearization and extend or mask the poly(A) sequence at the 3' end. It is not clear, whether this nonphysiological overhang affects the amount of protein produced intracellularly from such a construct.

RNA has several advantages over more traditional plasmid or viral approaches. Gene expression from an RNA source does not require transcription and the protein product is produced rapidly after the transfection. Further, since the RNA has to only gain access to the cytoplasm, rather than the nucleus, and therefore typical transfection methods result in an extremely high rate of transfection. In addition, plasmid based approaches require that the promoter driving the expression of the gene of interest be active in the cells under study.

In another aspect, the RNA construct is delivered into the cells by electroporation. See, e.g., the formulations and methodology of electroporation of nucleic acid constructs into mammalian cells as taught in US 2004/0014645, US 2005/0052630A1, US 2005/0070841 Al, US 2004/0059285A1, US 2004/0092907A1. The various parameters including electric field strength required for electroporation of any known cell type are generally known in the relevant research literature as well as numerous patents and applications in the field. See e.g., U.S. Pat. No. 6,678,556, U.S. Pat. No. 7,171,264, and U.S. Pat. No. 7,173,116. Apparatus for therapeutic application of electroporation are available commercially, e.g., the MedPulser™ DNA Electroporation Therapy System (Inovio/Genetronics, San Diego, Calif.), and are described in patents such as U.S. Pat.

No. 6,567,694; U.S. Pat. No. 6,516,223, U.S. Pat. No. 5,993,434, U.S. Pat. No. 6,181,964, U.S. Pat. No. 6,241,701, and U.S. Pat. No. 6,233,482; electroporation may also be used for transfection of cells in vitro as described e.g. in US20070128708A1. Electroporation may also be utilized to deliver nucleic acids into cells in vitro. Accordingly, electroporation-mediated administration into cells of nucleic acids including expression constructs utilizing any of the many available devices and electroporation systems known to those of skill in the art presents an exciting new means for delivering an RNA of interest to a target cell.

Sources of Cells

In one embodiment, cells are obtained from a subject. Non-limiting examples of subjects include humans, dogs, cats, mice, rats, pigs and transgenic species thereof. Preferably, the subject is a human. Cells can be obtained from a number of sources, including peripheral blood mononuclear cells, bone marrow, lymph node tissue, spleen tissue, umbilical cord, cancer cells and tumors. In certain embodiments, any number of cell lines available in the art, may be used. In certain embodiments, cells can be obtained from a unit of blood collected from a subject using any number of techniques known to the skilled artisan, such as Ficoll separation. In one embodiment, cells from the circulating blood of an individual are obtained by apheresis or leukapheresis. The apheresis product typically contains lymphocytes, including T cells, monocytes, granulocytes, B cells, other nucleated white blood cells, red blood cells, and platelets.

The cells collected by apheresis may be washed to remove the plasma fraction and to place the cells in an appropriate buffer or media, such as phosphate buffered saline (PBS) or wash solution lacks calcium and may lack magnesium or may lack many if not all divalent cations, for subsequent processing steps. After washing, the cells may be resuspended in a variety of biocompatible buffers, such as, for example, Ca-free, Mg-free PBS. Alternatively, the undesirable components of the apheresis sample may be removed and the cells directly resuspended in culture media.

In another embodiment, cells are isolated from peripheral blood. Alternatively, cells can be isolated from umbilical cord. In any event, a specific subpopulation of cells can be further isolated by positive or negative selection techniques.

Cells can also be frozen. While many freezing solutions and parameters are known in the art and will be useful in this context, in a non-limiting example, one method involves using PBS containing 20% DMSO and 8% human serum albumin, or other suitable cell freezing media. The cells are then frozen to -80°C at a rate of 1° per minute and stored in the vapor phase of a liquid nitrogen storage tank. Other methods of controlled freezing may be used as well as uncontrolled freezing immediately at -20°C or in liquid nitrogen.

Pharmaceutical compositions

Pharmaceutical compositions of the present invention may comprise the vector or modified cell as described herein, in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants ( e.g ., aluminum hydroxide); and preservatives. Compositions of the present invention are preferably formulated for intravenous administration.

Pharmaceutical compositions of the present invention may be administered in a manner appropriate to the disease to be treated (or prevented). The quantity and frequency of administration will be determined by such factors as the condition of the patient, and the type and severity of the patient’s disease, although appropriate dosages may be determined by clinical trials.

It can generally be stated that a pharmaceutical composition comprising the modified cells described herein may be administered at a dosage of 10 ⁴ to 10 ⁹ cells/kg body weight, in some instances 10 ⁵ to 10 ⁶ cells/kg body weight, including all integer values within those ranges. Compositions of the invention may also be administered multiple times at these dosages. The cells or vectors can be administered by using infusion techniques that are commonly known in immunotherapy (see, e.g., Rosenberg et ah, New Eng. J. of Med. 319: 1676, 1988). The optimal dosage and treatment regime for a particular patient can readily be determined by one skilled in the art of medicine by monitoring the patient for signs of disease and adjusting the treatment accordingly.

The administration of the modified cells or vectors of the invention may be carried out in any convenient manner known to those of skill in the art. The cells or vectors of the present invention may be administered to a subject by aerosol inhalation, injection, ingestion, transfusion, implantation or transplantation. The compositions described herein may be administered to a patient transarterially, subcutaneously, intradermally, intratumorally, intranodally, intramedullarly, intracystically intramuscularly, by intravenous (z.v.) injection, parenterally or intraperitoneally. In other instances, the cells of the invention are injected directly into a site of inflammation in the subject, a local disease site in the subject, a lymph node, an organ, a tumor, and the like.

It should be understood that the method and compositions that would be useful in the present invention are not limited to the particular formulations set forth in the examples. The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the cells, expansion and culture methods, and therapeutic methods of the invention, and are not intended to limit the scope of what the inventors regard as their invention.

The practice of the present invention employs, unless otherwise indicated, conventional techniques of molecular biology (including recombinant techniques), microbiology, cell biology, biochemistry and immunology, which are well within the purview of the skilled artisan. Such techniques are explained fully in the literature, such as, Sambrook et al. (2012 ) Molecular Cloning: A Laboratory Manual, fourth edition.

Cold Spring Harbor NY, Cold Spring Harbor Lab Press; Freshney, R.I. (2010) Culture of Animal Cells: A Manual of Basic Technique and Specialized Applications, sixth editionJo n Wiley & Sons; Ausubel et al. (2002) Short Protocols in Molecular Biology . John Wiley & Sons; Coligan et al. (2002) Current Protocols in Immunology. John Wiley & Sons. These techniques are applicable to the production of the polynucleotides and polypeptides of the invention, and, as such, may be considered in making and practicing the invention. Particularly useful techniques for particular embodiments will be discussed in the sections that follow.

EXPERIMENTAL EXAMPLES

The invention is now described with reference to the following Examples. These Examples are provided for the purpose of illustration only, and the invention is not limited to these Examples, but rather encompasses all variations that are evident as a result of the teachings provided herein.

The materials and methods employed in these experiments are now described.

APCM sgRNA library: The APCM library was designed by first selecting a small set of genes that elicit immune responses: B2M, B7-H2 (ICOSL), Calnexin, Calreticulin, CCL5, CD30L(TNFSF8), CD40L, CD70, CD80, CD83, CD86, CXCL10, CXCL3, CXCL9, Cystatin B, Cystatin C (x), ERAP1, ERp57(PDIA3), Flt3L, GITRL(TNF SF 18), IFNa4, IFNbl, IFNg, IL-2, LIGHT (TNFRSF14), NLRC5/CITA, OX40L/TNFSF4, Sec61al, Sec61b, Sec61g, TAPI, TAP2, Tapasin, T APB PR, TL1A (TNFSF15), and TNFSF9 (4-1BBL). SgRNA sequences were designed to target these genes in mice, yielding a library of 107 sgRNAs (SEQ ID NOs. 86-192) (Fig. 7). The sgRNAs were cloned into a viral vector and packaged.

CLC4 (CD80-Light-CXCL10-4-1BBL), CLC4G (CD80-Light-CXCL10-4-lBBL- GITRL) and CLC4I (CD80-Light-CXCL10-4-lBBL-IFNG): AAV-ORF-CLC4 was generated by concatenating ORFs of the CD80, Light, CXCL10, and 4-1BBL genes and expressing in an AAV vector (SEQ ID NO: 2). AAV-ORF-CLC4I was generated by concatenating ORFs of the CD80, Light, CXCL10, 4-1BBL, and IFNG genes expressing in an AAV vector (SEQ ID NO: 3). AAV-0-MAEGI-CLC4G was generated by cloning sgRNAs targeting the 5 genes into the AAV-CRISPRa vector (pGW059), and packaging into an AAV virus as a small pool: CD80-sg2 (NM_009855): TCCAGGCCTGTTCTGA GCAC (SEQ ID NO: 19), LIGHT (TNFRSF14)-sg2 (NM 019418): GAGGAGGT ACGT GAGGAAAG (SEQ ID NO: 20), CXCL10-sg3 (NM 021274): GCAATGCCCTCGGTT TACAG (SEQ ID NO: 21), TNFSF9 (4-lBBL)-sg2 (NM 009404): ACAGGGCCTGG ACAGGGAAG (SEQ ID NO: 22), GITRL(TNFSF18)-sgl (NM_183391): AGTGCTTAGCAGTGTTCCAA (SEQ ID NO: 23).

Table 1 : Vectors used in the invention pGW059 Lenti CRISPRa Screen Vector (SEQ ID NO: 1) ttaatgtagtcttatgcaatactcttgtagtcttgcaacatggtaacgatgagttagcaa catgccttacaaggagagaaaaagcacc gtgcatgccgattggtggaagtaaggtggtacgatcgtgccttattaggaaggcaacaga cgggtctgacatggattggacgaa ccactgaattgccgcattgcagagatattgtatttaagtgcctagctcgatacataaacg ggtctctctggttagaccagatctgagc ctgggagctctctggctaactagggaacccactgcttaagcctcaataaagcttgccttg agtgcttcaagtagtgtgtgcccgtct gttgtgtgactctggtaactagagatccctcagacccttttagtcagtgtggaaaatctc tagcagtggcgcccgaacagggactt gaaagcgaaagggaaaccagaggagctctctcgacgcaggactcggcttgctgaagcgcg cacggcaagaggcgaggggc ggcgactggtgagtacgccaaaaattttgactagcggaggctagaaggagagagatgggt gcgagagcgtcagtattaagcg ggggagaattagatcgcgatgggaaaaaattcggttaaggccagggggaaagaaaaaata taaattaaaacatatagtatgggc aagcagggagctagaacgattcgcagttaatcctggcctgttagaaacatcagaaggctg tagacaaatactgggacagctaca accatcccttcagacaggatcagaagaacttagatcattatataatacagtagcaaccct ctattgtgtgcatcaaaggatagagat aaaagacaccaaggaagctttagacaagatagaggaagagcaaaacaaaagtaagaccac cgcacagcaagcggccgctga tcttcagacctggaggaggagatatgagggacaattggagaagtgaattatataaatata aagtagtaaaaattgaaccattagga gtagcacccaccaaggcaaagagaagagtggtgcagagagaaaaaagagcagtgggaata ggagctttgttccttgggttctt gggagcagcaggaagcactatgggcgcagcgtcaatgacgctgacggtacaggccagaca attattgtctggtatagtgcagc agcagaacaatttgctgagggctattgaggcgcaacagcatctgttgcaactcacagtct ggggcatcaagcagctccaggcaa gaatcctggctgtggaaagatacctaaaggatcaacagctcctggggatttggggttgct ctggaaaactcatttgcaccactgct gtgccttggaatgctagttggagtaataaatctctggaacagatttggaatcacacgacc tggatggagtgggacagagaaattaa caattacacaagcttaatacactccttaattgaagaatcgcaaaaccagcaagaaaagaa tgaacaagaattattggaattagata aatgggcaagtttgtggaattggtttaacataacaaattggctgtggtatataaaattat tcataatgatagtaggaggcttggtaggt ttaagaatagtttttgctgtactttctatagtgaatagagttaggcagggatattcacca ttatcgtttcagacccacctcccaaccccg aggggaccctctagagatccgacgcgccatctctaggcccgcgccggccccctcgcacgg acttgtgggagaagctcggcta ctcccctgccccggttaatttgcatataatatttcctagtaactatagaggcttaatgtg cgataaaagacagataatctgttctttttaa tactagctacattttacatgataggcttggatttctataacttcgtatagcatacattat acgaagttataaacagcacaaaaggaaact caccctaactgtaaagtaattgtgtgttttgagactataaGtatcccttggagaaCCAcc tTGTTGgGAGACGggata cCGTCTCtgtttAagagctaAGCTGgccAACATGAGGATCACCCATGTCTGCAGggcCAG

CTtagcaagttaaaataaggctagtccgttatcaacttggccAACATGAGGATCACC CATGTCTGCAGg gccaagtggcaccgagtcggtgcTTTTTTGGATCCaagcttggcgtaactagatcttgag acaaatggcagtattcatc cacaattttaaaagaaaaggggggattggggggtacagtgcaggggaaagaatagtagac ataatagcaacagacatacaaac taaagaattacaaaaacaaattacaaaaattcaaaattttcgggtttattacagggacag cagagatccactttggcgccggctcga gggggcccgggtgcaaagatggataaagttttaaacagagaggaatctttgcagctaatg gaccttctaggtcttgaaaggagtg ggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccacagtccccgagaag ttggggggaggggtcggcaatt gatccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggc tccgcctttttcccgagggtgggg gagaaccgtatataagtgcagtagtcgccgtgaacgttctttttcgcaacgggtttgccg ccagaacacaggtaagtgccgtgtgt ggttcccgcgggcctggcctctttacgggttatggcccttgcgtgccttgaattacttcc actggctgcagtacgtgattcttgatcc cgagcttcgggttggaagtgggtgggagagttcgaggccttgcgcttaaggagccccttc gcctcgtgcttgagttgaggcctg gcctgggcgctggggccgccgcgtgcgaatctggtggcaccttcgcgcctgtctcgctgc tttcgataagtctctagccatttaaa atttttgatgacctgctgcgacgctttttttctggcaagatagtcttgtaaatgcgggcc aagatctgcacactggtatttcggtttttgg ggccgcgggcggcgacggggcccgtgcgtcccagcgcacatgttcggcgaggcggggcct gcgagcgcggccaccgaga atcggacgggggtagtctcaagctggccggcctgctctggtgcctggcctcgcgccgccg tgtatcgccccgccctgggcggc aaggctggcccggtcggcaccagttgcgtgagcggaaagatggccgcttcccggccctgc tgcagggagctcaaaatggag gacgcggcgctcgggagagcgggcgggtgagtcacccacacaaaggaaaagggcctttcc gtcctcagccgtcgcttcatgt gactccacggagtaccgggcgccgtccaggcacctcgattagttctcgagcttttggagt acgtcgtctttaggttggggggagg ggttttatgcgatggagtttccccacactgagtgggtggagactgaagttaggccagctt ggcacttgatgtaattctccttggaatt tgccctttttgagtttggatcttggttcattctcaagcctcagacagtggttcaaagttt ttttcttccatttcaggtgtcgtgacgtacgg ccaccatgaccgagtacaagcccacggtgcgcctcgccacccgcgacgacgtccccaggg ccgtacgcaccctcgccgccg cgttcgccgactaccccgccacgcgccacaccgtcgatccggaccgccacatcgagcggg tcaccgagctgcaagaactctt cctcacgcgcgtcgggctcgacatcggcaaggtgtgggtcgcggacgacggcgccgccgt ggcggtctggaccacgccgg agagcgtcgaagcgggggcggtgttcgccgagatcggcccgcgcatggccgagttgagcg gttcccggctggccgcgcagc aacagatggaaggcctcctggcgccgcaccggcccaaggagcccgcgtggttcctggcca ccgtcggagtctcgcccgacc accagggcaagggtctgggcagcgccgtcgtgctccccggagtggaggcggccgagcgcg ccggggtgcccgccttcctg gagacctccgcgccccgcaacctccccttctacgagcggctcggcttcaccgtcaccgcc gacgtcgaggtgcccgaaggac cgcgcacctggtgcatgacccgcaagcccggtgccGGATCCGGCGCAACAAACTTCTCTC TGCTG

AAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggaagacgccaaaaacata aag aaaggcccggcgccattctatccgctggaagatggaaccgctggagagcaactgcataag gctatgaagagatacgccctggt tcctggaacaattgcttttacagatgcacatatcgaggtggacatcacttacgctgagta cttcgaaatgtccgttcggttggcagaa gctatgaaacgatatgggctgaatacaaatcacagaatcgtcgtatgcagtgaaaactct cttcaattctttatgccggtgttgggcg cgttatttatcggagttgcagttgcgcccgcgaacgacatttataatgaacgtgaattgc tcaacagtatgggcatttcgcagcctac cgtggtgttcgtttccaaaaaggggttgcaaaaaattttgaacgtgcaaaaaaagctccc aatcatccaaaaaattattatcatggat tctaaaacggattaccagggatttcagtcgatgtacacgttcgtcacatctcatctacct cccggttttaatgaatacgattttgtgcca gagtccttcgatagggacaagacaattgcactgatcatgaactcctctggatctactggt ctgcctaaaggtgtcgctctgcctcat agaactgcctgcgtgagattctcgcatgccagagatcctatttttggcaatcaaatcatt ccggatactgcgattttaagtgttgttcc attccatcacggttttggaatgtttactacactcggatatttgatatgtggatttcgagt cgtcttaatgtatagatttgaagaGgagct gtttctgaggagccttcaggattacaagattcaaagtgcgctgctggtgccaaccctatt ctccttcttcgccaaaagcactctgatt gacaaatacgatttatctaatttacacgaaattgcttctggtggcgctcccctctctaag gaagtcggggaagcggttgccaagag gttccatctgccaggtatcaggcaaggatatgggctcactgagactacatcagctattct gattacacccgagggggatgataaac cgggcgcggtcggtaaagttgttccattttttgaagcgaaggttgtggatctggataccg ggaaaacgctgggcgttaatcaaag aggcgaactgtgtgtgagaggtcctatgattatgtccggttatgtaaacaatccggaagc gaccaacgccttgattgacaaggatg gatggctacattctggagacatagcttactgggacgaagacgaacacttcttcatcgttg accgcctgaagtctctgattaagtaca aaggctatcaggtggctcccgctgaattggaatccatcttgctccaacaccccaacatct tcgacgcaggtgtcgcaggtcttccc gacgatgacgccggtgaacttcccgccgccgttgttgttttggagcacggaaagacgatg acggaaaaagagatcgtggattac gtcgccagtcaagtaacaaccgcgaaaaagttgcgcggaggagttgtgtttgtggacgaa gtaccgaaaggtcttaccggaaaa ctcgacgcaagaaaaatcagagagatcctcataaaggccaagaagggcggaaagatcgcc gtgTAAgaattcacgcgttaa gtcgacaatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactat gttgctccttttacgctatgtggatacgctg ctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttgt ataaatcctggttgctgtctctttatgaggagttg tggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccact ggttggggcattgccaccacctgtc agctcctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccg cctgccttgcccgctgctggacaggg gctcggctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttcct tggctgctcgcctgtgttgccacctgga ttctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttcctt cccgcggcctgctgccggctctgcgg cctcttccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctcc ccgcgtcgactttaagaccaatgactt acaaggcagctgtagatcttagccactttttaaaagaaaaggggggactggaagggctaa ttcactcccaacgaagacaagatct gctttttgcttgtactgggtctctctggttagaccagatctgagcctgggagctctctgg ctaactagggaacccactgcttaagcct caataaagcttgccttgagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggt aactagagatccctcagacccttttagtc agtgtggaaaatctctagcagtacgtatagtagttcatgtcatcttattattcagtattt ataacttgcaaagaaatgaatatcagagag tgagaggaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaa tttcacaaataaagcatttttttcactgca ttctagttgtggtttgtccaaactcatcaatgtatcttatcatgtctggctctagctatc ccgcccctaactccgcccatcccgccccta actccgcccagttccgcccattctccgccccatggctgactaattttttttatttatgca gaggccgaggccgcctcggcctctgag ctattccagaagtagtgaggaggcttttttggaggcctagggacgtacccaattcgccct atagtgagtcgtattacgcgcgctca ctggccgtcgttttacaacgtcgtgactgggaaaaccctggcgttacccaacttaatcgc cttgcagcacatccccctttcgccag ctggcgtaatagcgaagaggcccgcaccgatcgcccttcccaacagttgcgcagcctgaa tggcgaatgggacgcgccctgta gcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgcca gcgccctagcgcccgctcctttc gctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgg gggctccctttagggttccgatttagtgct ttacggcacctcgaccccaaaaaacttgattagggtgatggttcacgtagtgggccatcg ccctgatagacggtttttcgccctttg acgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaac cctatctcggtctattcttttgatttataag ggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacg cgaattttaacaaaatattaacgcttacaa tttaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaat acattcaaatatgtatccgctcatgagacaa taaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttc cgtgtcgcccttattcccttttttgcggcat tttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatc agttgggtgcacgagtgggttacatcg aactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaa tgatgagcacttttaaagttctgctatgt ggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactat tctcagaatgacttggttgagtactc accagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgc cataaccatgagtgataacactgcg gccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaac atgggggatcatgtaactcgccttga tcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcc tgtagcaatggcaacaacgttgc gcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactgga tggaggcggataaagttgcaggacc acttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggtga gcgtgggtctcgcggtatcattgcagc actggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggc aactatggatgaacgaaatagaca gatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactc atatatactttagattgatttaaaacttca tttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatccc ttaacgtgagttttcgttccactgagcgtca gaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgc tgcttgcaaacaaaaaaaccaccgctac cagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggct tcagcagagcgcagataccaaatact gttcttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctaca tacctcgctctgctaatcctgttaccagt ggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttacc ggataaggcgcagcggtcgggctg aacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagata cctacagcgtgagctatgagaa agcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcgga acaggagagcgcacgaggg agcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgac ttgagcgtcgatttttgtgatgctcgtca ggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttt tgctggccttttgctcacatgttcttt cctgcgttatcccctgattctgtggataaccgtattaccgcctttgagtgagctgatacc gctcgccgcagccgaacgaccgagcg cagcgagtcagtgagcgaggaagcggaagagcgcccaatacgcaaaccgcctctccccgc gcgttggccgattcattaatgc agctggcacgacaggtttcccgactggaaagcgggcagtgagcgcaacgcaattaatgtg agttagctcactcattaggcaccc caggctttacactttatgcttccggctcgtatgttgtgtggaattgtgagcggataacaa tttcacacaggaaacagctatgaccatg attacgccaagcgcgcaattaaccctcactaaagggaacaaaagctggagctgcaagc pGW066 AAV-ORF-CLC4 (SEQ ID NO: 2)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgctgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaAgtgctgggctggccactc agggctggtttctcctgagactgc atcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggaga agctgatacaagatcaacgatctc accaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtg gacctctgttatgggagacacgac ttggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagc ccggttactactatgtgtactccaaa gtgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccat ggactatacaagcgcacatccc gctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaaca gctcccgagtctggtgggacag cagcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcc tggaaaccgcctggtcagacca cgtgacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGC A AC AAACTTCTCTCT

GCTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgaacccaagtgctgc cg tcattttctgcctcatcctgctgggtctgagtgggactcaagggatccctctcgcaagga cggtccgctgcaactgcatccatatcg atgacgggccagtgagaatgagggccatagggaagcttgaaatcatccctgcgagcctat cctgcccacgtgttgagatcattg ccacgatgaaaaagaatgatgagcagagatgtctgaatccggaatctaagaccatcaaga atttaatgaaagcgtttagccaaaa aaggtctaaaagggctcctgctagcagcggtacccagtgcaccaactacgccctgctgaa gctggccggcgatgtggagagc aaccccgggcccatggaccagcacacacttgatgtggaggataccgcggatgccagacat ccagcaggtacttcgtgcccctc ggatgcggcgctcctcagagataccgggctcctcgcggacgctgcgctcctctcagatac tgtgcgccccacaaatgccgcgc tccccacggatgctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgc ctgcactgaacttctgttcccgcca cccaaagctctatggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcc tatcttcacccgcaccgagcctcggcca gcgctcacaatcaccacctcgcccaacctgggtacccgagagaataatgcagaccaggtc acccctgtttcccacattggctgc cccaacactacacaacagggctctcctgtgttcgccaagctactggctaaaaaccaagca tcgttgtgcaatacaactctgaactg gcacagccaagatggagctgggagctcatacctatctcaaggtctgaggtacgaagaaga caaaaaggagttggtggtagaca gtcccgggctctactacgtatttttggaactgaagctcagtccaacattcacaaacacag gccacaaggtgcagggctgggtctct cttgttttgcaagcaaagcctcaggtagatgactttgacaacttggccctgacagtggaa ctgttcccttgctccatggagaacaag ttagtggaccgttcctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtg ggtctgagggcttatctgcatggagc ccaggatgcatacagagactgggagctgtcttatcccaacaccaccagctttggactctt tcttAtgaaacccgacaacccatgg gaatgaGaattcacgcgttaagtcgacaatcaacctctggattacaaaatttgtgaaaga ttgactggtattcttaactatgttgctcc ttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttcccgtat ggctttcattttctcctccttgtataaatcctggt tgctgtctctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtgtgcactg tgtttgctgacgcaacccccactggtt ggggcattgccaccacctgtcagctcctttccgggactttcgctttccccctccctattg ccacggcggaactcatcgccgcctgc cttgcccgctgctggacaggggctcggctgttgggcactgacaattccgtggtgttgtcg gggaaatcatcgtcctttccttggctg ctcgcctgtgttgccacctggattctgcgcgggacgtccttctgctacgtcccttcggcc ctcaatccagcggaccttccttcccgc ggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagacgagtcgg atctccctttgggccgcctccccgc gtcgactttaagaccaatgacGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCT C

TCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGC

CCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGC

AGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACAC

CGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGC

GGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTA

GCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTT

CCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTT

ACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGG

CCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTT

AATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTA

TTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGA

GCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAA

TTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAG

CCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCC

GGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAG

AGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATAC

GCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGT

GGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAAT

ACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAAT

AATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATT

CCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTG

AAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAAC

TGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTT

CCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTAT

TGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGAC

TTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAG

TAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAA

CTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACA

AC AT GGGGGATC ATGT AACTCGCCTT GATCGTT GGGAACCGGAGCTGAAT GA

AGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACA

ACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACA

ATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCG

GCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGG

AAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATC

GT AGTT ATCTACAC G AC GGGG AGT C AGGC A AC TAT GG AT G A AC G A A AT AG AC

AGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCA AGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAG

GATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTG

AGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCT

TGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACC

GCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGA

AGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAG

CCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGC

TCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTA

C C GGGTT GG AC T C A AG AC GAT AGT T AC C GG AT A AGGC GC AGC GGT C GGGC T G

AACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAA

CTGAGAT ACCT AC AGCGT GAGCT AT GAGAAAGCGCC ACGCTTCCCGAAGGGA

GAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCA

CGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTT

CGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAG

CCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCT

GGCCTTTTGCTCACATGT pGW067 AAV-ORF-CLC4I (SEQ ID NO: 3)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgTtgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctgggctggccactc agggctggtttctcctgagactgca tcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggagaa gctgatacaagatcaacgatctca ccaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtgg acctctgttatgggagacacgact tggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagcc cggttactactatgtgtactccaaag tgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccatg gactatacaagcgcacatcccg ctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaacag ctcccgagtctggtgggacagca gcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcctg gaaaccgcctggtcagaccacgt gacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGCAACAAAC TTCTCTCTG

CTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgaacccaagtgctgcc gtc attttctgcctcatcctgctgggtctgagtgggactcaagggatccctctcgcaaggacg gtccgctgcaactgcatccatatcgat gacgggccagtgagaatgagggccatagggaagcttgaaatcatccctgcgagcctatcc tgcccacgtgttgagatcattgcc acgatgaaaaagaatgatgagcagagatgtctgaatccggaatctaagaccatcaagaat ttaatgaaagcgtttagccaaaaaa ggtctaaaagggctcctgctagcagcggtacccagtgcaccaactacgccctgctgaagc tggccggcgatgtggagagcaa ccccgggcccatggaccagcacacacttgatgtggaggataccgcggatgccagacatcc agcaggtacttcgtgcccctcgg atgcggcgctcctcagagataccgggctcctcgcggacgctgcgctcctctcagatactg tgcgccccacaaatgccgcgctcc ccacggatgctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctg cactgaacttctgttcccgccaccc aaagctctatggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctat cttcacccgcaccgagcctcggccagcg ctcacaatcaccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacc cctgtttcccacattggctgcccca acactacacaacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgt tgtgcaatacaactctgaactggcac agccaagatggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaa aaggagttggtggtagacagtcc cgggctctactacgtatttttggaactgaagctcagtccaacattcacaaacacaggcca caaggtgcagggctgggtctctcttg ttttgcaagcaaagcctcaggtagatgactttgacaacttggccctgacagtggaactgt tcccttgctccatggagaacaagttag tggaccgttcctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtc tgagggcttatctgcatggagccca ggatgcatacagagactgggagctgtcttatcccaacaccaccagctttggactctttct tgtgaaacccgacaacccatgggaa ggcatatgcggtaccgtgaagcagaccctgaacttcgatctgctgaagctggccggcgat gtggagagcaaccccgggcccat gaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgtta ctgccacggcacagtcattgaaagccta gaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctcttc ttggatatctggaggaactggcaaaag gatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttgaa gtcttgaaagacaatcaggccatcagc aacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggcg aaaaaggatgcattcatgagtattgc caagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgagt ggtccaccagctgttgccggaatc cagcctcaggaagcggaaaaggagtcgctgctgaGAATTCAATAAAAGATCTTTATTTTC ATTAG

ATCTGTGT GTT GGTTTTTTGT GT GCGGCCGC AGGAACCCCT AGT GAT GGAGTT

GGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAG

GTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGC

GCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGC

GGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGG

CGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTT

GCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACG

TTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCG

ATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTT

CACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAG

TCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTC

TATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTG

GTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATA

TTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGC

ATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGG

GCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAG

CTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAG

GGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCT

TAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTT

ATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGAT

AAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCG

TGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCA

GAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGG

GTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCC

GAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGT ATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATT CTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGA T GGC AT G AC AGT A AG AG A AT TAT GC AGT GC T GC CAT A AC CAT G AGT GAT A AC ACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCG _{CTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCG} GAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAG CAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCT TCCCGGC AAC AATT AAT AGACTGGAT GGAGGCGGAT AAAGTT GC AGGACC AC TTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCC GGT GAGCGTGGAAGCCGCGGT ATC ATT GC AGC ACTGGGGCC AGAT GGT AAGC CCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGA ACGAAAT AGAC AGATCGCTGAGAT AGGT GCCTC ACTGATT AAGC ATTGGT AA CTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTT TAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAAT CCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCA AAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAA AAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAAC TCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTC TTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCT ACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAA GTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAG CGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGA CCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCT T C C C G A AGGG AG A A AGGC GG AC AGGT ATC C GGT AAGC GGC AGGGT C GG A AC AGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGT CCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCA GGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW029 Adenovirus One Vector dSpCas9 (SEQ ID NO: 4)

CTACAGGGCGCGTCCATTCGCCATTCAGGATCGAATTAATTCTTAATTAACAT

CAT C A AT AAT AT AC CTT Attttggattgaagccaatatgataatgagggggtggagtttgtgacgtggcgcgggg cgtgggaacggggcgggtgacgtagtagtgtggcggaagtgtgatgttgcaagtgtggcg gaacacatgtaagcgacggatgt ggcaaaagtgacgtttttggtgtgcgccggtgtacacaggaagtgacaattttcgcgcgg ttttaggcggatgttgtagtaaatttg ggcgtaaccgagtaagatttggccattttcgcgggaaaactgaataagaggaagtgaaat ctgaataattttgtgttactcatagcg cgtaaTACTggtaccgcggccgcctcgagtCTAGAAAGAgggcctatttcccatgattcc ttcatatttgcatatacga tacaaggctgttagagagataattagaattaatttgactgtaaacacaaagatattagta caaaatacgtgacgtagaaagtaataat ttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccg taacttgaaagtatttcgatttcttggctttatat atcttGTGGAAAGGACGAAACACCggaagagcgagctcttctgttttagagctaggccAA CATGAGG

ATCACCCATGTCTGCAGggcctagcaagttaaaataaggctagtccgttatcaactt ggccAACATGAGG

ATCACCCATGTCTGCAGggccaagtggcaccgagtcggtgcTTTTTTGctagcGTCA GTGGGC

AGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAAT

T G A AC GGGT GC C T AG AG A AGGT GGC GC GGGGT A A AC T GGG A A AGT GAT GT C

GTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGC

AGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGC

TGAAGCTTCGAGGGGCTCGCATCTCTCCTTCACGCGCCCGCCGCCCTACCTGA

GGCCGCCATCCACGCCGGTTGAGTCGCGTTCTGCCGCCTCCCGCCTGTGGTGC

CTCCTGAACTGCGTCCGCCGTCTAGGTAAGTTTAAAGCTCAGGTCGAGACCGG

GCCTTTGTCCGGCGCTCCCTTGGAGCCTACCTAGACTCAGCCGGCTCTCCACG

CTTTGCCTGACCCTGCTTGCTCAACTCTACGTCTTTGTTTCGTTTTCTGTTCTGC GCCGTTACAGATCCAAGctcGTACGgccaccATGAAAAGGCCGGCGGCCACGAAA

AAGGCCGGCCAGGCAAAAAAGAAAAAGGACAAGAAGTACAGCATCGGCCTG

GCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGTACAAGG

TGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAA

GAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCC

ACCCGGCTGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAACCGG

ATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGCCAAGGTGGACGACA

GCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCA

CGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAG

AAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCGACA

AGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGTTCCGG

GGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGGACA

AGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCC

CATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGC

AAGAGCAGACGGCTGGAAAATCTGATCGCCCAGCTGCCCGGCGAGAAGAAG

AATGGCCTGTTCGGCAACCTGATTGCCCTGAGCCTGGGCCTGACCCCCAACTT

C AAGAGC AACTTCGACCTGGCCGAGGAT GCC AAACTGC AGCTGAGC AAGGAC

ACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACG

CCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGAC

ATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGCCTCTATGA

TCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGT

GCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAG

AACGGCTACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTCTACA

AGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCACCGAGGAACTGCTCGT

GAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGG

CAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGG

CAGGAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGA

TCCTGACCTTCCGCATCCCCTACTACGTGGGCCCTCTGGCCAGGGGAAACAGC

AGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGGAACT

TCGAGGA AGTGGT GGAC AAGGGCGCTTCCGCCC AGAGCTTC ATCGAGCGGAT

GACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGC

CTGCTGTACGAGTACTTCACCGTGTATAACGAGCTGACCAAAGTGAAATACG

TGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAGAAAAAGG

CCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCT

GAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCC

GGCGTGGAAGATCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCTGA

AAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAACGAGGACATTCT

GGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAG

GAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGC

TGAAGCGGCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGATCA

ACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTGGATTTCCTGAAGTC

CGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTG

ACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCC

TGCACGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCAT

CCTGC AGAC AGTGA AGGTGGT GGACGAGCTCGT GAAAGT GAT GGGCCGGC AC

AAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCAG

A AGGG AC AGA AG A AC AGC CGC GAG AG A AT GA AGC GGAT C GA AGAGGGC AT C

AAAGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACCCAG

CTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAATGGGCGGGATATGT

ACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGACCA CATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTG

ACCAGAAGCGACAAGGCCCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAG

GT C GT G A AG A AG AT G A AG A AC TACT GGC GGC AGC T GC T G A AC GC C A AGC T G A

TTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGGCCTGAG

CGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAG

AT C AC A A AGC AC GT GGC AC AGATCC T GG ACTC CC GGAT G A AC AC T A AGT ACG

ACGAGAAT GAC AAGCTGATCCGGGAAGT GAAAGT GAT C ACCCTGAAGTCC AA

GCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGTGCGCGAGATCA

ACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGC

CCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTAC

AAGGTGTACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAATCGGC

AAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCATGAACTTTTTCAAGAC

CGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGAGACA

AACGGCGAAACCGGGGAGATCGT GTGGGAT AAGGGCCGGGATTTT GCC ACCG

TGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGT

GCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAGCGAT

AAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCG

AC AGCCCC ACCGT GGCCT ATTCTGT GCTGGTGGT GGCC AAAGT GGAAAAGGG

C A AGT C C A AGA A ACTGA AGAGT GT GA A AGAGCTGCTGGGGAT C AC CAT CAT G

GAAAGAAGCAGCTTCGAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCT

ACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACTCCCTGTT

CGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAG

AAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGG

CCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACA

GCTGTTT GTGGAAC AGC AC AAGC ACT ACCTGGACGAGAT C ATCGAGC AGAT C

AGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGC

TGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGA

ATATCATCCACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAG

TACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCAAAGAGGTGC

TGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGAT

CGACCTGTCTCAGCTGGGAGGCGACagcgctGGAggacctaagaaaaagaggaaggt gggatccG

GACGGGCTGACGCATTGGACGATTTTGATCTGGATATGCTGGGAAGTGACGC

CCTCGATGATTTTGACCTTGACATGCTTGGTTCGGATGCCCTTGATGACTTTGA

CCTCGACATGCTCGGCAGTGACGCCCTTGATGATTTCGACCTGGACATGCTGA

TTAACtgtacaggaagcggagccactaacttctccctgttgaaacaagcaggggatg tcgaagagaatcccgggccaGC

CACCATGGCTTCAAACTTTACTCAGTTCGTGCTCGTGGACAATGGTGGGACAG

GGGATGTGACAGTGGCTCCTTCTAATTTCGCTAATGGGGTGGCAGAGTGGATC

AGCTCCAACTCACGGAGCCAGGCCTACAAGGTGACATGCAGCGTCAGGCAGT

CT AGT GCCC AGAAGAGA AAGT AT ACC ATC AAGGT GGAGGTCCCC AAAGT GGC

TACCCAGACAGTGGGCGGAGTCGAACTGCCTGTCGCCGCTTGGAGGTCCTAC

CTGAACATGGAGCTCACTATCCCAATTTTCGCTACCAATTCTGACTGTGAACT

CATCGTGAAGGCAATGCAGGGGCTCCTCAAAGACGGTAATCCTATCCCTTCC

GCC ATCGCCGCTAACTCAGGTATCTAC AGC GCTGGAGGAGGTGG AAGC GGAG

GAGGAGGA AGCGGAGGAGGAGGT AGC GGACC T A AGA A A A AGAGGA AGGT G

GCGGCCGCTGGATCCCCTTCAGGGCAGATCAGCAACCAGGCCCTGGCTCTGG

CCCCTAGCTCCGCTCCAGTGCTGGCCCAGACTATGGTGCCCTCTAGTGCTATG

GTGCCTCTGGCCCAGCCACCTGCTCCAGCCCCTGTGCTGACCCCAGGACCACC

CCAGTCACTGAGCGCTCCAGTGCCCAAGTCTACACAGGCCGGCGAGGGGACT

CTGAGTGAAGCTCTGCTGCACCTGCAGTTCGACGCTGATGAGGACCTGGGAG

CTCTGCTGGGGAACAGCACCGATCCCGGAGTGTTCACAGATCTGGCCTCCGTG GACAACTCTGAGTTTCAGCAGCTGCTGAATCAGGGCGTGTCCATGTCTCATAG

TACAGCCGAACCAATGCTGATGGAGTACCCCGAAGCCATTACCCGGCTGGTG

ACCGGCAGCCAGCGGCCCCCCGACCCCGCTCCAACTCCCCTGGGAACCAGCG

GCCTGCCTAATGGGCTGTCCGGAGATGAAGACTTCTCAAGCATCGCTGATATG

GACTTTAGTGCCCTGCTGTCACAGATTTCCTCTAGTGGGCAGGGAGGAGGTGG

AAGCGGCTTCAGCGTGGACACCAGTGCCCTGCTGGACCTGTTCAGCCCCTCG

GTGACCGTGCCCGACATGAGCCTGCCTGACCTTGACAGCAGCCTGGCCAGTA

TCCAAGAGCTCCTGTCTCCCCAGGAGCCCCCCAGGCCTCCCGAGGCAGAGAA

CAGCAGCCCGGATTCAGGGAAGCAGCTGGTGCACTACACAGCGCAGCCGCTG

TTCCTGCTGGACCCCGGCTCCGTGGACACCGGGAGCAACGACCTGCCGGTGC

TGTTTGAGCTGGGAGAGGGCTCCTACTTCTCCGAAGGGGACGGCTTCGCCGA

GGACCCCACCATCTCCCTGCTGACAGGCTCGGAGCCTCCCAAAGCCAAGGAC

CCCACTGTCTCCTAAGAATTCgatatcaagcttAATAAAAGATCTTTATTTTCATTA G

ATCTGTGT GTT GGTTTTTTGT GT agatctGGGCGT GGTtaagggtgggaaagaatatataaggtgg gggtcttatgtagttttgtatctgttttgcagcagccgccgccgccatgagcaccaactc gtttgatggaagcattgtgagctcatatt tgacaacgcgcatgcccccatgggccggggtgcgtcagaatgtgatgggctccagcattg atggtcgccccgtcctgcccgca aactctactaccttgacctacgagaccgtgtctggaacgccgttggagactgcagcctcc gccgccgcttcagccgctgcagcc accgcccgcgggattgtgactgactttgctttcctgagcccgcttgcaagcagtgcagct tcccgttcatccgcccgcgatgacaa gttgacggctcttttggcacaattggattctttgacccgggaacttaatgtcgtttctca gcagctgttggatctgcgccagcaggttt ctgccctgaaggcttcctcccctcccaatgcggtttaaaacataaataaaaaaccagact ctgtttggatttggatcaagcaagtgt cttgctgtctttatttaggggttttgcgcgcgcggtaggcccgggaccagcggtctcggt cgttgagggtcctgtgtattttttccag gacgtggtaaaggtgactctggatgttcagatacatgggcataagcccgtctctggggtg gaggtagcaccactgcagagcttc atgctgcggggtggtgttgtagatgatccagtcgtagcaggagcgctgggcgtggtgcct aaaaatgtctttcagtagcaagctg attgccaggggcaggcccttggtgtaagtgtttacaaagcggttaagctgggatgggtgc atacgtggggatatgagatgcatctt ggactgtatttttaggttggctatgttcccagccatatccctccggggattcatgttgtg cagaaccaccagcacagtgtatccggtg cacttgggaaatttgtcatgtagcttagaaggaaatgcgtggaagaacttggagacgccc ttgtgacctccaagattttccatgcat tcgtccataatgatggcaatgggcccacgggcggcggcctgggcgaagatatttctggga tcactaacgtcatagttgtgttcca ggatgagatcgtcataggccatttttacaaagcgcgggcggagggtgccagactgcggta taatggttccatccggcccaggg gcgtagttaccctcacagatttgcatttcccacgctttgagttcagatggggggatcatg tctacctgcggggcgatgaagaaaac ggtttccggggtaggggagatcagctgggaagaaagcaggttcctgagcagctgcgactt accgcagccggtgggcccgtaa atcacacctattaccgggtgcaactggtagttaagagagctgcagctgccgtcatccctg agcaggggggccacttcgttaagca tgtccctgactcgcatgttttccctgaccaaatccgccagaaggcgctcgccgcccagcg atagcagttcttgcaaggaagcaaa gtttttcaacggtttgagaccgtccgccgtaggcatgcttttgagcgtttgaccaagcag ttccaggcggtcccacagctcggtca cctgctctacggcatctcgatccagcatatctcctcgtttcgcgggttggggcggctttc gctgtacggcagtagtcggtgctcgtc cagacgggccagggtcatgtctttccacgggcgcagggtcctcgtcagcgtagtctgggt cacggtgaaggggtgcgctccgg gctgcgcgctggccagggtgcgcttgaggctggtcctgctggtgctgaagcgctgccggt cttcgccctgcgcgtcggccagg tagcatttgaccatggtgtcatagtccagcccctccgcggcgtggcccttggcgcgcagc ttgcccttggaggaggcgccgcac gaggggcagtgcagacttttgagggcgtagagcttgggcgcgagaaataccgattccggg gagtaggcatccgcgccgcag gccccgcagacggtctcgcattccacgagccaggtgagctctggccgttcggggtcaaaa accaggtttcccccatgctttttga tgcgtttcttacctctggtttccatgagccggtgtccacgctcggtgacgaaaaggctgt ccgtgtccccgtatacagactnnngtt taaacgaattcnnntataaaatgcaaggtgctgctcaaaaaatcaggcaaagcctcgcgc aaaaaagaaagcacatcgtagtca tgctcatgcagataaaggcaggtaagctccggaaccaccacagaaaaagacaccattttt ctctcaaacatgtctgcgggtttctg cataaacacaaaataaaataacaaaaaaacatttaaacattagaagcctgtcttacaaca ggaaaaacaacccttataagcataag acggactacggccatgccggcgtgaccgtaaaaaaactggtcaccgtgattaaaaagcac caccgacagctcctcggtcatgtc cggagtcataatgtaagactcggtaaacacatcaggttgattcatcggtcagtgctaaaa agcgaccgaaatagcccgggggaa tacatacccgcaggcgtagagacaacattacagcccccataggaggtataacaaaattaa taggagagaaaaacacataaaca cctgaaaaaccctcctgcctaggcaaaatagcaccctcccgctccagaacaacatacagc gcttcacagcggcagcctaacagt cagccttaccagtaaaaaagaaaacctattaaaaaaacaccactcgacacggcaccagct caatcagtcacagtgtaaaaaagg gccaagtgcagagcgagtatatataggactaaaaaatgacgtaacggttaaagtccacaa aaaacacccagaaaaccgcacgc gaacctacgcccagaaacgaaagccaaaaaacccacaacttcctcaaatcgtcacttccg ttttcccacgttacgtaacttcccatt ttaagaaaactacaattcccaacacatacaagttactccgccctaaaacctacgtcaccc gccccgttcccacgccccgcgccac gtcacaaactccaccccctcattatcatattggcttcaatccaaaataaggtatattatt gatgattaattaattaaggatcccggtgtg aaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttacgcttcctcgc tcactgactcgctgcgctcggtcgt tcggctgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatc aggggataacgcaggaaagaaca tgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttt tccataggctccgcccccctgac gagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaaga taccaggcgtttccccctggaag ctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttct cccttcgggaagcgtggcgctttctcat agctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtg cacgaaccccccgttcagcccgacc gctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgc cactggcagcagccactggtaacag gattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaacta cggctacactagaaggacagtattt ggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatcc ggcaaacaaaccaccgctggtagcg gtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatc ctttgatcttttctacggggtctgacgct cagtggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttc acctagatccttttaaattaaaaatgaa gttttaaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaa tcagtgaggcacctatctcagcgatctgtc tatttcgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagg gcttaccatctggccccagtgctgcaat gataccgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccgg aagggccgagcgcagaagtggt cctgcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagt agttcgccagttaatagtttgcgcaacgt tgttgnnnnnnaaaaaggatcttcacctagatccttttcacgtagaaagccagtccgcag aaacggtgctgaccccggatgaat gtcagctactgggctatctggacaagggaaaacgcaagcgcaaagagaaagcaggtagct tgcagtgggcttacatggcgata gctagactgggcggttttatggacagcaagcgaaccggaattgccagctggggcgccctc tggtaaggttgggaagccctgca aagtaaactggatggctttctcgccgccaaggatctgatggcgcaggggatcaagctctg atcaagagacaggatgaggatcgt ttcgcatgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggc tattcggctatgactgggcacaaca gacaatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttct ttttgtcaagaccgacctgtccggt gccctgaatgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgtt ccttgcgcagctgtgctcgacgt tgtcactgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcct gtcatctcaccttgctcctgccga gaaagtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctg cccattcgaccaccaagcgaaacat cgcatcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggac gaggagcatcaggggctcgcgc cagccgaactgttcgccaggctcaaggcgagcatgcccgacggcgaggatctcgtcgtga cccatggcgatgcctgcttgccg aatatcatggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtg gcggaccgctatcaggacatagcgtt ggctacccgtgatattgctgaggagcttggcggcgaatgggctgaccgcttcctcgtgct ttacggtatcgccgctcccgattcgc agcgcatcgccttctatcgccttcttgacgagttcttctgaattttgttaaaatttttgt taaatcagctcattttttaaccaataggccgaa atcggcaacatcccttataaatcaaaagaatagaccgcgatagggttgagtgttgttcca gtttggaacaagagtccactattaaag aacgtggactccaacgtcaaagggcgaaaaaccgtctatcagggcgatggcccactacgt gaaccatcacccaaatcaagttttt tgcggtcgaggtgccgtaaagctctaaatcggaaccctaaagggagcccccgatttagag cttgacggggaaagccggcgaa cgtggcgagaaaggaagggaagaaagcgaaaggagcgggcgctagggcgctggcaagtgt agcggtcacgctgcgcgta accaccacacccgcgcgcttaatgcgccg pGW035 Adenovirus One Vector dSpCas9 (SEQ ID NO: 5)

CTACAGGGCGCGTCCATTCGCCATTCAGGATCGAATTAATTCTTAATTAACAT

CAT C A AT A AT AT AC CTT Attttggattgaagccaatatgataatgagggggtggagtttgtgacgtggcgcgggg cgtgggaacggggcgggtgacgtagtagtgtggcggaagtgtgatgttgcaagtgtggcg gaacacatgtaagcgacggatgt ggcaaaagtgacgtttttggtgtgcgccggtgtacacaggaagtgacaattttcgcgcgg ttttaggcggatgttgtagtaaatttg ggcgtaaccgagtaagatttggccattttcgcgggaaaactgaataagaggaagtgaaat ctgaataattttgtgttactcatagcg cgtaaTACTggtaccgcggccgcctcgagtCTAGAAAGAgggcctatttcccatgattcc ttcatatttgcatatacga tacaaggctgttagagagataattagaattaatttgactgtaaacacaaagatattagta caaaatacgtgacgtagaaagtaataat ttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgcttaccg taacttgaaagtatttcgatttcttggctttatat atcttGTGGAAAGGACGAAACACCggaagagcgagctcttctgttttagagctaggccAA CATGAGG

ATCACCCATGTCTGCAGggcctagcaagttaaaataaggctagtccgttatcaactt ggccAACATGAGG

ATCACCCATGTCTGCAGggccaagtggcaccgagtcggtgcTTTTTTGctagcGTCA GTGGGC

AGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGCAAT T G A AC GGGT GC C T AG AG A AGGT GGC GC GGGGT A A AC T GGG A A AGT GAT GT C

GTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAGTGC

AGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACACAGC

TGAAGCTTCGAGGGGCTCGCATCTCTCCTTCACGCGCCCGCCGCCCTACCTGA

GGCCGCCATCCACGCCGGTTGAGTCGCGTTCTGCCGCCTCCCGCCTGTGGTGC

CTCCTGAACTGCGTCCGCCGTCTAGGTAAGTTTAAAGCTCAGGTCGAGACCGG

GCCTTTGTCCGGCGCTCCCTTGGAGCCTACCTAGACTCAGCCGGCTCTCCACG

CTTTGCCTGACCCTGCTTGCTCAACTCTACGTCTTTGTTTCGTTTTCTGTTCTGC

GCCGTTACAGATCCAAGctcGTACGgccaccATGAAAAGGCCGGCGGCCACGAAA

AAGGCCGGCCAGGCAAAAAAGAAAAAGGACAAGAAGTACAGCATCGGCCTG

GCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGTACAAGG

TGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCATCAA

GAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAGGCC

ACCCGGCTGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAACCGG

ATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGCCAAGGTGGACGACA

GCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAAGCA

CGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCACGAG

AAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCGACA

AGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGTTCCGG

GGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGGACA

AGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAACCC

CATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTGAGC

AAGAGCAGACGGCTGGAAAATCTGATCGCCCAGCTGCCCGGCGAGAAGAAG

AATGGCCTGTTCGGCAACCTGATTGCCCTGAGCCTGGGCCTGACCCCCAACTT

C AAGAGC AACTTCGACCTGGCCGAGGAT GCC AAACTGC AGCTGAGC AAGGAC

ACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAGTACG

CCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAGCGAC

ATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGCCTCTATGA

TCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTCTCGT

GCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAGCAAG

AACGGCTACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTCTACA

AGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCACCGAGGAACTGCTCGT

GAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAACGG

CAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGGCGG

CAGGAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGAAGA

TCCTGACCTTCCGCATCCCCTACTACGTGGGCCCTCTGGCCAGGGGAAACAGC

AGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGGAACT

TCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCGGAT

GACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCACAGC

CTGCTGTACGAGTACTTCACCGTGTATAACGAGCTGACCAAAGTGAAATACG

TGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAGAAAAAGG

CCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCAGCT

GAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGGAAATCTCC

GGCGTGGAAGATCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCTGA

AAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAACGAGGACATTCT

GGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCGAG

GAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCAGC

TGAAGCGGCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGATCA

ACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTGGATTTCCTGAAGTC

CGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCCTG ACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAGCC

TGCACGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGCAT

CCTGC AGAC AGTGA AGGTGGT GGACGAGCTCGT GAAAGT GAT GGGCCGGC AC

AAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCCAG

A AGGG AC AGA AG A AC AGC CGC GAG AG A AT GA AGC GGAT C GA AGAGGGC AT C

AAAGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACCCAG

CTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAATGGGCGGGATATGT

ACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGACCA

CATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTGCTG

ACCAGAAGCGACAAGGCCCGGGGCAAGAGCGACAACGTGCCCTCCGAAGAG

GT C GT G A AG A AG AT G A AG A AC TACT GGC GGC AGC T GC T G A AC GC C A AGC T G A

TTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGGCGGCCTGAG

CGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCGGCAG

ATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAGTACG

ACGAGAAT GAC AAGCTGATCCGGGAAGT GAAAGT GAT C ACCCTGAAGTCC AA

GCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGTGCGCGAGATCA

ACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAACCGC

CCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGACTAC

AAGGTGTACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAATCGGC

AAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCATGAACTTTTTCAAGAC

CGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGAGACA

AACGGCGAAACCGGGGAGATCGT GTGGGAT AAGGGCCGGGATTTT GCC ACCG

TGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCGAGGT

GCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAGCGAT

AAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTACGGCGGCTTCG

AC AGCCCC ACCGT GGCCT ATTCTGT GCTGGTGGT GGCC AAAGT GGAAAAGGG

C A AGT C C A AGA A ACTGA AGAGT GT GA A AGAGCTGCTGGGGAT C AC CAT CAT G

GAAAGAAGCAGCTTCGAGAAGAATCCCATCGACTTTCTGGAAGCCAAGGGCT

ACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACTCCCTGTT

CGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAACTGCAG

AAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTACCTGG

CCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAGAAACA

GCTGTTT GTGGAAC AGC AC AAGC ACT ACCTGGACGAGAT C ATCGAGC AGAT C

AGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAAAGTGC

TGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCCGAGA

ATATCATCCACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTTCAAG

TACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCAAAGAGGTGC

TGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACACGGAT

CGACCTGTCTCAGCTGGGAGGCGACagcgctGGAGGAGGTGGAAGCGGAGGAG

GAGGAAGCGGAGGAGGAGGTAGCggacctaagaaaaagaggaaggtggcggccgctg gatccGGA

CGGGCTGACGCATTGGACGATTTTGATCTGGATATGCTGGGAAGTGACGCCCT

CGATGATTTTGACCTTGACATGCTTGGTTCGGATGCCCTTGATGACTTTGACCT

CGACATGCTCGGCAGTGACGCCCTTGATGATTTCGACCTGGACATGCTGATTA

ACtgtacaggaagcggagccactaacttctccctgttgaaacaagcaggggatgtcg aagagaatcccgggccaGCCAC

CATGGCTTCAAACTTTACTCAGTTCGTGCTCGTGGACAATGGTGGGACAGGGG

ATGTGACAGTGGCTCCTTCTAATTTCGCTAATGGGGTGGCAGAGTGGATCAGC

TCCAACTCACGGAGCCAGGCCTACAAGGTGACATGCAGCGTCAGGCAGTCTA

GTGCCCAGAAGAGAAAGTATACCATCAAGGTGGAGGTCCCCAAAGTGGCTAC

CCAGACAGTGGGCGGAGTCGAACTGCCTGTCGCCGCTTGGAGGTCCTACCTG

AACATGGAGCTCACTATCCCAATTTTCGCTACCAATTCTGACTGTGAACTCAT CGTGAAGGCAATGCAGGGGCTCCTCAAAGACGGTAATCCTATCCCTTCCGCC

ATCGCCGCTAACTCAGGTATCTACAGCGCTGGAGGAGGTGGAAGCGGAGGAG

GAGGAAGCGGAGGAGGAGGTAGCGGACCTAAGAAAAAGAGGAAGGTGGCG

GCCGCTGGATCCCCTTCAGGGCAGATCAGCAACCAGGCCCTGGCTCTGGCCC

CTAGCTCCGCTCCAGTGCTGGCCCAGACTATGGTGCCCTCTAGTGCTATGGTG

CCTCTGGCCCAGCCACCTGCTCCAGCCCCTGTGCTGACCCCAGGACCACCCCA

GTCACTGAGCGCTCCAGTGCCCAAGTCTACACAGGCCGGCGAGGGGACTCTG

AGTGAAGCTCTGCTGCACCTGCAGTTCGACGCTGATGAGGACCTGGGAGCTC

TGCTGGGGAACAGCACCGATCCCGGAGTGTTCACAGATCTGGCCTCCGTGGA

CAACTCTGAGTTTCAGCAGCTGCTGAATCAGGGCGTGTCCATGTCTCATAGTA

CAGCCGAACCAATGCTGATGGAGTACCCCGAAGCCATTACCCGGCTGGTGAC

CGGCAGCCAGCGGCCCCCCGACCCCGCTCCAACTCCCCTGGGAACCAGCGGC

CTGCCTAATGGGCTGTCCGGAGATGAAGACTTCTCAAGCATCGCTGATATGG

ACTTTAGTGCCCTGCTGTCACAGATTTCCTCTAGTGGGCAGGGAGGAGGTGGA

AGCGGCTTCAGCGTGGACACCAGTGCCCTGCTGGACCTGTTCAGCCCCTCGGT

GACCGTGCCCGACATGAGCCTGCCTGACCTTGACAGCAGCCTGGCCAGTATC

CAAGAGCTCCTGTCTCCCCAGGAGCCCCCCAGGCCTCCCGAGGCAGAGAACA

GCAGCCCGGATTCAGGGAAGCAGCTGGTGCACTACACAGCGCAGCCGCTGTT

CCTGCTGGACCCCGGCTCCGTGGACACCGGGAGCAACGACCTGCCGGTGCTG

TTTGAGCTGGGAGAGGGCTCCTACTTCTCCGAAGGGGACGGCTTCGCCGAGG

ACCCCACCATCTCCCTGCTGACAGGCTCGGAGCCTCCCAAAGCCAAGGACCC

CACTGTCTCCTAAGAATTCgatatcaagcttAATAAAAGATCTTTATTTTCATTAGA T

CTGTGTGTTGGTTTTTTGTGTagatctGGGCGTGGTtaagggtgggaaagaatatat aaggtgggggt cttatgtagttttgtatctgttttgcagcagccgccgccgccatgagcaccaactcgttt gatggaagcattgtgagctcatatttgac aacgcgcatgcccccatgggccggggtgcgtcagaatgtgatgggctccagcattgatgg tcgccccgtcctgcccgcaaact ctactaccttgacctacgagaccgtgtctggaacgccgttggagactgcagcctccgccg ccgcttcagccgctgcagccaccg cccgcgggattgtgactgactttgctttcctgagcccgcttgcaagcagtgcagcttccc gttcatccgcccgcgatgacaagttg acggctcttttggcacaattggattctttgacccgggaacttaatgtcgtttctcagcag ctgttggatctgcgccagcaggtttctgc cctgaaggcttcctcccctcccaatgcggtttaaaacataaataaaaaaccagactctgt ttggatttggatcaagcaagtgtcttgc tgtctttatttaggggttttgcgcgcgcggtaggcccgggaccagcggtctcggtcgttg agggtcctgtgtattttttccaggacgt ggtaaaggtgactctggatgttcagatacatgggcataagcccgtctctggggtggaggt agcaccactgcagagcttcatgctg cggggtggtgttgtagatgatccagtcgtagcaggagcgctgggcgtggtgcctaaaaat gtctttcagtagcaagctgattgcc aggggcaggcccttggtgtaagtgtttacaaagcggttaagctgggatgggtgcatacgt ggggatatgagatgcatcttggact gtatttttaggttggctatgttcccagccatatccctccggggattcatgttgtgcagaa ccaccagcacagtgtatccggtgcactt gggaaatttgtcatgtagcttagaaggaaatgcgtggaagaacttggagacgcccttgtg acctccaagattttccatgcattcgtc cataatgatggcaatgggcccacgggcggcggcctgggcgaagatatttctgggatcact aacgtcatagttgtgttccaggatg agatcgtcataggccatttttacaaagcgcgggcggagggtgccagactgcggtataatg gttccatccggcccaggggcgtag ttaccctcacagatttgcatttcccacgctttgagttcagatggggggatcatgtctacc tgcggggcgatgaagaaaacggtttcc ggggtaggggagatcagctgggaagaaagcaggttcctgagcagctgcgacttaccgcag ccggtgggcccgtaaatcacac ctattaccgggtgcaactggtagttaagagagctgcagctgccgtcatccctgagcaggg gggccacttcgttaagcatgtccct gactcgcatgttttccctgaccaaatccgccagaaggcgctcgccgcccagcgatagcag ttcttgcaaggaagcaaagtttttc aacggtttgagaccgtccgccgtaggcatgcttttgagcgtttgaccaagcagttccagg cggtcccacagctcggtcacctgct ctacggcatctcgatccagcatatctcctcgtttcgcgggttggggcggctttcgctgta cggcagtagtcggtgctcgtccagac gggccagggtcatgtctttccacgggcgcagggtcctcgtcagcgtagtctgggtcacgg tgaaggggtgcgctccgggctgc gcgctggccagggtgcgcttgaggctggtcctgctggtgctgaagcgctgccggtcttcg ccctgcgcgtcggccaggtagca tttgaccatggtgtcatagtccagcccctccgcggcgtggcccttggcgcgcagcttgcc cttggaggaggcgccgcacgagg ggcagtgcagacttttgagggcgtagagcttgggcgcgagaaataccgattccggggagt aggcatccgcgccgcaggcccc gcagacggtctcgcattccacgagccaggtgagctctggccgttcggggtcaaaaaccag gtttcccccatgctttttgatgcgtt tcttacctctggtttccatgagccggtgtccacgctcggtgacgaaaaggctgtccgtgt ccccgtatacagactnnngtttaaac gaattcnnntataaaatgcaaggtgctgctcaaaaaatcaggcaaagcctcgcgcaaaaa agaaagcacatcgtagtcatgctc atgcagataaaggcaggtaagctccggaaccaccacagaaaaagacaccatttttctctc aaacatgtctgcgggtttctgcataa acacaaaataaaataacaaaaaaacatttaaacattagaagcctgtcttacaacaggaaa aacaacccttataagcataagacgg actacggccatgccggcgtgaccgtaaaaaaactggtcaccgtgattaaaaagcaccacc gacagctcctcggtcatgtccgga gtcataatgtaagactcggtaaacacatcaggttgattcatcggtcagtgctaaaaagcg accgaaatagcccgggggaatacat acccgcaggcgtagagacaacattacagcccccataggaggtataacaaaattaatagga gagaaaaacacataaacacctga aaaaccctcctgcctaggcaaaatagcaccctcccgctccagaacaacatacagcgcttc acagcggcagcctaacagtcagc cttaccagtaaaaaagaaaacctattaaaaaaacaccactcgacacggcaccagctcaat cagtcacagtgtaaaaaagggcca agtgcagagcgagtatatataggactaaaaaatgacgtaacggttaaagtccacaaaaaa cacccagaaaaccgcacgcgaac ctacgcccagaaacgaaagccaaaaaacccacaacttcctcaaatcgtcacttccgtttt cccacgttacgtaacttcccattttaag aaaactacaattcccaacacatacaagttactccgccctaaaacctacgtcacccgcccc gttcccacgccccgcgccacgtcac aaactccaccccctcattatcatattggcttcaatccaaaataaggtatattattgatga ttaattaattaaggatcccggtgtgaaata ccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttacgcttcctcgctcact gactcgctgcgctcggtcgttcggc tgcggcgagcggtatcagctcactcaaaggcggtaatacggttatccacagaatcagggg ataacgcaggaaagaacatgtga gcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttccat aggctccgcccccctgacgagc atcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacaggactataaagatacc aggcgtttccccctggaagctcc ctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgtccgcctttctccct tcgggaagcgtggcgctttctcatagct cacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagctgggctgtgtgcacg aaccccccgttcagcccgaccgctg cgccttatccggtaactatcgtcttgagtccaacccggtaagacacgacttatcgccact ggcagcagccactggtaacaggatta gcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaactacggct acactagaaggacagtatttggtat ctgcgctctgctgaagccagttaccttcggaaaaagagttggtagctcttgatccggcaa acaaaccaccgctggtagcggtggt ttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttg atcttttctacggggtctgacgctcagt ggaacgaaaactcacgttaagggattttggtcatgagattatcaaaaaggatcttcacct agatccttttaaattaaaaatgaagtttt aaatcaatctaaagtatatatgagtaaacttggtctgacagttaccaatgcttaatcagt gaggcacctatctcagcgatctgtctattt cgttcatccatagttgcctgactccccgtcgtgtagataactacgatacgggagggctta ccatctggccccagtgctgcaatgat accgcgagacccacgctcaccggctccagatttatcagcaataaaccagccagccggaag ggccgagcgcagaagtggtcct gcaactttatccgcctccatccagtctattaattgttgccgggaagctagagtaagtagt tcgccagttaatagtttgcgcaacgttgt tgnnnnnnaaaaaggatcttcacctagatccttttcacgtagaaagccagtccgcagaaa cggtgctgaccccggatgaatgtc agctactgggctatctggacaagggaaaacgcaagcgcaaagagaaagcaggtagcttgc agtgggcttacatggcgatagct agactgggcggttttatggacagcaagcgaaccggaattgccagctggggcgccctctgg taaggttgggaagccctgcaaag taaactggatggctttctcgccgccaaggatctgatggcgcaggggatcaagctctgatc aagagacaggatgaggatcgtttcg catgattgaacaagatggattgcacgcaggttctccggccgcttgggtggagaggctatt cggctatgactgggcacaacagac aatcggctgctctgatgccgccgtgttccggctgtcagcgcaggggcgcccggttctttt tgtcaagaccgacctgtccggtgcc ctgaatgaactgcaagacgaggcagcgcggctatcgtggctggccacgacgggcgttcct tgcgcagctgtgctcgacgttgtc actgaagcgggaagggactggctgctattgggcgaagtgccggggcaggatctcctgtca tctcaccttgctcctgccgagaaa gtatccatcatggctgatgcaatgcggcggctgcatacgcttgatccggctacctgccca ttcgaccaccaagcgaaacatcgca tcgagcgagcacgtactcggatggaagccggtcttgtcgatcaggatgatctggacgagg agcatcaggggctcgcgccagc cgaactgttcgccaggctcaaggcgagcatgcccgacggcgaggatctcgtcgtgaccca tggcgatgcctgcttgccgaatat catggtggaaaatggccgcttttctggattcatcgactgtggccggctgggtgtggcgga ccgctatcaggacatagcgttggct acccgtgatattgctgaggagcttggcggcgaatgggctgaccgcttcctcgtgctttac ggtatcgccgctcccgattcgcagc gcatcgccttctatcgccttcttgacgagttcttctgaattttgttaaaatttttgttaa atcagctcattttttaaccaataggccgaaatc ggcaacatcccttataaatcaaaagaatagaccgcgatagggttgagtgttgttccagtt tggaacaagagtccactattaaagaa cgtggactccaacgtcaaagggcgaaaaaccgtctatcagggcgatggcccactacgtga accatcacccaaatcaagttttttg cggtcgaggtgccgtaaagctctaaatcggaaccctaaagggagcccccgatttagagct tgacggggaaagccggcgaacg tggcgagaaaggaagggaagaaagcgaaaggagcgggcgctagggcgctggcaagtgtag cggtcacgctgcgcgtaacc accacacccgcgcgcttaatgcgccg pGW063 Adenovirus One Vector dSpCas9 (SEQ ID NO: 6)

AATTAATTAAGCTAGCATCATCAATAATATACCTTATTTTGGATTGAAGCCAA T ATGAT AATGAGGGGGT GGAGTTT GT GACGT GGCGCGGGGCGTGGGAACGGG GCGGGTGACGTAGTAGTGTGGCGGAAGTGTGATGTTGCAAGTGTGGCGGAAC ACATGTAAGCGACGGATGTGGCAAAAGTGACGTTTTTGGTGTGCGCCGGTGT

ACACAGGAAGTGACAATTTTCGCGCGGTTTTAGGCGGATGTTGTAGTAAATTT

GGGCGTAACCGAGTAAGATTTGGCCATTTTCGCGGGAAAACTGAATAAGAGG

AAGTGAAATCTGAATAATTTTGTGTTACTCATAGCGCGTAATATTTGTCTAGG

GAGATCCGGTACcgcggccgcctcgagtCTAGAAAGAgggcctatttcccatgattc cttcatatttgcatata cgatacaaggctgttagagagataattagaattaatttgactgtaaacacaaagatatta gtacaaaatacgtgacgtagaaagtaa taatttcttgggtagtttgcagttttaaaattatgttttaaaatggactatcatatgctt accgtaacttgaaagtatttcgatttcttggcttt atatatcttGTGGAAAGGACGAAACACCggaagagcgagctcttctgttttagagctagg ccAACATGA

GGAT C AC CC AT GT C T GC AGggcctagcaagttaaaataaggctagtccgttatcaacttggcc A AC AT GA

GGATCACCCATGTCTGCAGggccaagtggcaccgagtcggtgcTTTTTTGctagcGT CAGTGG

GCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCGGC

AATTGAACGGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTGAT

GTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATAAG

TGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAACAC

AGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTCACGCGCCCGCCGCCCTACC

TGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTCTGCCGCCTCCCGCCTGTGG

TGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGTTTAAAGCTCAGGTCGAGAC

CGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACCTAGACTCAGCCGGCTCTCC

ACGCTTTGCCTGACCCTGCTTGCTCAACTCTACGTCTTTGTTTCGTTTTCTGTTC

TGCGCCGTTACAGATCCAAGctcGTACGgccaccATGAAAAGGCCGGCGGCCACG

AAAAAGGCCGGCCAGGCAAAAAAGAAAAAGGACAAGAAGTACAGCATCGGC

CTGGCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGTACA

AGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACAGCAT

CAAGAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGCCGAG

GCCACCCGGCTGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAAGAAC

CGGATCTGCTATCTGCAAGAGATCTTCAGCAACGAGATGGCCAAGGTGGACG

ACAGCTTCTTCCACAGACTGGAAGAGTCCTTCCTGGTGGAAGAGGATAAGAA

GCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTACCAC

GAGAAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGCACCG

ACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCAAGTTC

CGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGACGTGG

ACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAGGAAAA

CCCCATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCAGACTG

AGCAAGAGCAGACGGCTGGAAAATCTGATCGCCCAGCTGCCCGGCGAGAAG

AAGAATGGCCTGTTCGGCAACCTGATTGCCCTGAGCCTGGGCCTGACCCCCA

ACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGAGCAA

GGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGACCAG

TACGCCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGCTGAG

CGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGCCTCT

ATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAAGCTC

TCGTGCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACCAGAG

CAAGAACGGCTACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAGAGTTC

TACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCACCGAGGAACTGC

TCGTGAAGCTGAACAGAGAGGACCTGCTGCGGAAGCAGCGGACCTTCGACAA

CGGCAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTCTGCGG

CGGCAGGAAGATTTTTACCCATTCCTGAAGGACAACCGGGAAAAGATCGAGA

AGATCCTGACCTTCCGCATCCCCTACTACGTGGGCCCTCTGGCCAGGGGAAAC

AGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCCCCTGGA

ACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCATCGAGCG

GATGACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCCAAGCAC AGCCTGCTGTACGAGTACTTCACCGTGTATAACGAGCTGACCAAAGTGAAAT

ACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCAGAAAA

AGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTGAAGCA

GCTGAAAGAGGACT ACTTC AAGAAAATCGAGT GCTTCGACTCCGTGGA AATC

TCCGGCGTGGAAGATCGGTTCAACGCCTCCCTGGGCACATACCACGATCTGCT

GAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAACGAGGACATT

CTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGATGATCG

AGGAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGATGAAGCA

GCTGAAGCGGCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAAGCTGAT

CAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTGGATTTCCTGAAG

TCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACGACAGCC

TGACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGGCGATAG

CCTGCACGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAGAAGGGC

AT C C T GC AG AC AGT G A AGGT GGT GG AC G AGC T C GT G A A AGT GAT GGGC C GGC

ACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGACCACCC

AGAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAGAGGGC

ATCAAAGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGGAAAACACC

CAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAATGGGCGGGATA

TGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGATGTGGA

CCACATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACAAGGTG

CTGACCAGAAGCGACAAGGCCCGGGGCAAGAGCGACAACGTGCCCTCCGAA

GAGGTCGTGAAGA AGATGAAGAACT ACTGGCGGC AGCTGCTGAACGCC A AG

CTGATT ACCC AGAGAAAGTTCGAC AATCTGACC A AGGCCGAGAGAGGCGGCC

TGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAAACCCG

GCAGATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACACTAAG

TACGACGAGAATGACAAGCTGATCCGGGAAGTGAAAGTGATCACCCTGAAGT

CCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGTGCGCGAG

ATCAACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGTGGGAA

CCGCCCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTACGGCGA

CTACAAGGTGTACGACGTGCGGAAGATGATCGCCAAGAGCGAGCAGGAAAT

CGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCATGAACTTTTTCA

AGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTGATCGA

GACAAACGGCGAAACCGGGGAGATCGTGTGGGATAAGGGCCGGGATTTTGCC

ACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAAAGACCG

AGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAGGAACAG

C GAT A AGC T GAT C GC C AG A A AG A AGG AC T GGG AC C C T A AG A AGT AC GGC GG

CTTCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAGTGGAAA

AGGGC A AGTCC A AGA A ACTGA AGAGT GT GA A AGAGC T GC T GGGGAT C ACC A

T CAT GGAAAGAAGC AGCTTCGAGAAGAATCCC ATCGACTTTCTGGAAGCC AA

GGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAAGTACTCC

CTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCCGGCGAAC

TGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACTTCCTGTA

CCTGGCCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAATGAGCAG

AAACAGCTGTTTGTGGAACAGCACAAGCACTACCTGGACGAGATCATCGAGC

AGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATCTGGACAA

AGTGCTGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGAGCAGGCC

GAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAGCCCCTGCCGCCTT

CAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCACCAAAGAG

GTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTACGAGACAC

GGATCGACCTGTCTCAGCTGGGAGGCGACagcgctGGAGGAGGTGGAAGCGGAG GAGGAGGAAGCGGAGGAGGAGGTAGCggacctaagaaaaagaggaaggtggcggccgctg GAT

CCCCTTCAGGGCAGATCAGCAACCAGGCCCTGGCTCTGGCCCCTAGCTCCGCT

CCAGTGCTGGCCCAGACTATGGTGCCCTCTAGTGCTATGGTGCCTCTGGCCCA

GCCACCTGCTCCAGCCCCTGTGCTGACCCCAGGACCACCCCAGTCACTGAGC

GCTCCAGTGCCCAAGTCTACACAGGCCGGCGAGGGGACTCTGAGTGAAGCTC

T GCTGC ACCTGC AGTTCGACGCTGATGAGGACCTGGGAGCTCTGCTGGGGAA

CAGCACCGATCCCGGAGTGTTCACAGATCTGGCCTCCGTGGACAACTCTGAGT

TTCAGCAGCTGCTGAATCAGGGCGTGTCCATGTCTCATAGTACAGCCGAACCA

ATGCTGATGGAGTACCCCGAAGCCATTACCCGGCTGGTGACCGGCAGCCAGC

GGCCCCCCGACCCCGCTCCAACTCCCCTGGGAACCAGCGGCCTGCCTAATGG

GCTGTCCGGAGATGAAGACTTCTCAAGCATCGCTGATATGGACTTTAGTGCCC

T GCTGTC AC AGATTTCCTCT AGTGGGC AGGGAGGAGGT GGAAGCGGCTTC AG

CGTGGACACCAGTGCCCTGCTGGACCTGTTCAGCCCCTCGGTGACCGTGCCCG

ACATGAGCCTGCCTGACCTTGACAGCAGCCTGGCCAGTATCCAAGAGCTCCT

GTCTCCCCAGGAGCCCCCCAGGCCTCCCGAGGCAGAGAACAGCAGCCCGGAT

TCAGGGAAGCAGCTGGTGCACTACACAGCGCAGCCGCTGTTCCTGCTGGACC

CCGGCTCCGTGGACACCGGGAGCAACGACCTGCCGGTGCTGTTTGAGCTGGG

AGAGGGCTCCTACTTCTCCGAAGGGGACGGCTTCGCCGAGGACCCCACCATC

TCCCTGCTGACAGGCTCGGAGCCTCCCAAAGCCAAGGACCCCACTGTCTCCTA

AGAATTCCTGCAGCCCGGGGGATCCACTAGTTCTAGAGCGGCCGCCACCGCG

GGGAGATCCAGACATGATAAGATACATTGATGAGTTTGGACAAACCACAACT

AGAATGCAGTGAAAAAAATGCTTTATTTGTGAAATTTGTGATGCTATTGCTTT

ATTTGTAACCATTATAAGCTGCAATAAACAAGTTAACAACAACAATTGCATTC

ATTTT AT GTTTC AGGTTC AGGGGGAGGT GT GGGAGGTTTTTT AAAGC A AGT AA

AACCTCTACAAATGTGGTATGGCTGATTATGATCCCGGCTGCCTCGCGCGTTT

CGGTGATGACGGTGAAAACCTCTTGACACATGCAGCTCCCGGAGACGGTCAC

AGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGCGTCA

GCGGGTGTTGGCGGGTGTCGGGGCGCAGCCATGAGGTCGACTCTAGTCCCCG

CGGTGGCAGATCTGGAAGGTGCTGAGGTACGATGAGACCCGCACCAGGTGCA

GACCCTGCGAGTGTGGCGGTAAACATATTAGGAACCAGCCTGTGATGCTGGA

TGTGACCGAGGAGCTGAGGCCCGATCACTTGGTGCTGGCCTGCACCCGCGCT

GAGTTT GGCTCT AGCGAT GAAGAT AC AGATT GAGGT ACTGAAAT GT GTGGGC

GTGGCTTAAGGGTGGGAAAGAATATATAAGGTGGGGGTCTTATGTAGTTTTG

TATCTGTTTTGCAGCAGCCGCCGCCGCCATGAGCACCAACTCGTTTGATGGAA

GCATTGTGAGCTCATATTTGACAACGCGCATGCCCCCATGGGCCGGGGTGCGT

CAGAATGTGATGGGCTCCAGCATTGATGGTCGCCCCGTCCTGCCCGCAAACTC

TACTACCTTGACCTACGAGACCGTGTCTGGAACGCCGTTGGAGACTGCAGCCT

CCGCCGCCGCTTCAGCCGCTGCAGCCACCGCCCGCGGGATTGTGACTGACTTT

GCTTTCCTGAGCCCGCTTGCAAGCAGTGCAGCTTCCCGTTCATCCGCCCGCGA

TGACAAGTTGACGGCTCTTTTGGCACAATTGGATTCTTTGACCCGGGAACTTA

ATGTCGTTTCTCAGCAGCTGTTGGATCTGCGCCAGCAGGTTTCTGCCCTGAAG

GCTTCCTCCCCTCCCAATGCGGTTTAAAACATAAATAAAAAACCAGACTCTGT

TTGGATTTGGATCAAGCAAGTGTCTTGCTGTCTTTATTTAGGGGTTTTGCGCGC

GCGGTAGGCCCGGGACCAGCGGTCTCGGTCGTTGAGGGTCCTGTGTATTTTTT

CCAGGACGTGGTAAAGGTGACTCTGGATGTTCAGATACATGGGCATAAGCCC

GTCTCTGGGGT GGAGGT AGC ACC ACTGC AGAGCTT CAT GCTGC GGGGTGGTG

TTGTAGATGATCCAGTCGTAGCAGGAGCGCTGGGCGTGGTGCCTAAAAATGT

CTTTCAGTAGCAAGCTGATTGCCAGGGGCAGGCCCTTGGTGTAAGTGTTTACA

A AGC GGT T A AGC T GGG AT GGGT GC AT AC GT GGGG AT AT GAG AT GC AT C T T GG

ACTGTATTTTTAGGTTGGCTATGTTCCCAGCCATATCCCTCCGGGGATTCATGT TGTGCAGAACCACCAGCACAGTGTATCCGGTGCACTTGGGAAATTTGTCATGT

AGCTTAGAAGGAAATGCGTGGAAGAACTTGGAGACGCCCTTGTGACCTCCAA

GATTTTCCATGCATTCGTCCATAATGATGGCAATGGGCCCACGGGCGGCGGCC

T GGGCGA AGAT ATTTCTGGGAT C ACT AACGT CAT AGTTGT GTTCC AGGAT GAG

ATCGTCATAGGCCATTTTTACAAAGCGCGGGCGGAGGGTGCCAGACTGCGGT

ATAATGGTTCCATCCGGCCCAGGGGCGTAGTTACCCTCACAGATTTGCATTTC

CCACGCTTTGAGTTCAGATGGGGGGATCATGTCTACCTGCGGGGCGATGAAG

AAAACGGTTTCCGGGGTAGGGGAGATCAGCTGGGAAGAAAGCAGGTTCCTGA

GCAGCTGCGACTTACCGCAGCCGGTGGGCCCGTAAATCACACCTATTACCGG

GTGCAACTGGTAGTTAAGAGAGCTGCAGCTGCCGTCATCCCTGAGCAGGGGG

GCCACTTCGTTAAGCATGTCCCTGACTCGCATGTTTTCCCTGACCAAATCCGC

CAGAAGGCGCTCGCCGCCCAGCGATAGCAGTTCTTGCAAGGAAGCAAAGTTT

TTCAACGGTTTGAGACCGTCCGCCGTAGGCATGCTTTTGAGCGTTTGACCAAG

CAGTTCCAGGCGGTCCCACAGCTCGGTCACCTGCTCTACGGCATCTCGATCCA

GCATATCTCCTCGTTTCGCGGGTTGGGGCGGCTTTCGCTGTACGGCAGTAGTC

GGTGCTCGTCCAGACGGGCCAGGGTCATGTCTTTCCACGGGCGCAGGGTCCTC

GTCAGCGTAGTCTGGGTCACGGTGAAGGGGTGCGCTCCGGGCTGCGCGCTGG

CCAGGGTGCGCTTGAGGCTGGTCCTGCTGGTGCTGAAGCGCTGCCGGTCTTCG

CCCTGCGCGTCGGCCAGGTAGCATTTGACCATGGTGTCATAGTCCAGCCCCTC

CGCGGCGTGGCCCTTGGCGCGCAGCTTGCCCTTGGAGGAGGCGCCGCACGAG

GGGCAGTGCAGACTTTTGAGGGCGTAGAGCTTGGGCGCGAGAAATACCGATT

CCGGGGAGTAGGCATCCGCGCCGCAGGCCCCGCAGACGGTCTCGCATTCCAC

GAGCCAGGTGAGCTCTGGCCGTTCGGGGTCAAAAACCAGGTTTCCCCCATGC

TTTTTGATGCGTTTCTTACCTCTGGTTTCCATGAGCCGGTGTCCACGCTCGGTG

ACGAAAAGGCTGTCCGTGTCCCCGTATACAGACTTGAGAGGCCTGTCCTCGA

CCGATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGG

GCATGACTATCGTCGCCGCACTTATGACTGTCTTCTTTATCATGCAACTCGTA

GGACAGGTGCCGGCAGCGCTCTGGGTCATTTTCGGCGAGGACCGCTTTCGCTG

GAGCGCGACGATGATCGGCCTGTCGCTTGCGGTATTCGGAATCTTGCACGCCC

TCGCTCAAGCCTTCGTCACTGGTCCCGCCACCAAACGTTTCGGCGAGAAGCA

GGCCATTATCGCCGGCATGGCGGCCGACGCGCTGGGCTACGTCTTGCTGGCGT

TCGCGACGCGAGGCTGGATGGCCTTCCCCATTATGATTCTTCTCGCTTCCGGC

GGCATCGGGATGCCCGCGTTGCAGGCCATGCTGTCCAGGCAGGTAGATGACG

ACCATCAGGGACAGCTTCAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGG

CCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAA

AATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACC

AGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCG

CTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAT

AGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGG

CTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACT

ATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGC

C AC T GGT A AC AGGATT AGC AG AGCGAGGT AT GT AGGCGGT GCT AC AGAGTT C

TTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTG

CGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCG

GC A AAC AAACC ACCGCTGGT AGC GGT GGTTTTTTT GTTT GC AAGC AGC AGATT

ACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGT

CTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATT

ATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAAT

CAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATC

AGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTG ACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCA

GTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGC

AATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTA

TCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTC

GCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTGCAGGCATCGTGGTGT

CACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGG

CGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCC

TCCGATCGTTGT C AGAAGT AAGTT GGCCGC AGT GTT ATC ACTC AT GGTT AT GG

CAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGA

CTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAG

TTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTT

TAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGAT

CTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGAT

CTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGG

CAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTC

ATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATG

AGC GGAT AC AT ATTTGA AT GT ATTT AGA A A A AT A A AC A A AT AGGGGTTCCGC

GCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATG

ACATTAACCTATAAAAATAGGCGTATCACGAGGCCCTTTCGTCTTCAAGAA pAdEasy-1 Adenovirus Packaging or Helper (SEQ ID NO: 7) attgtgagctcatatttgacaacgcgcatgcccccatgggccggggtgcgtcagaatgtg atgggctccagcattgatggtcgcc ccgtcctgcccgcaaactctactaccttgacctacgagaccgtgtctggaacgccgttgg agactgcagcctccgccgccgcttc agccgctgcagccaccgcccgcgggattgtgactgactttgctttcctgagcccgcttgc aagcagtgcagcttcccgttcatcc gcccgcgatgacaagttgacggctcttttggcacaattggattctttgacccgggaactt aatgtcgtttctcagcagctgttggatc tgcgccagcaggtttctgccctgaaggcttcctcccctcccaatgcggtttaaaacataa ataaaaaaccagactctgtttggatttg gatcaagcaagtgtcttgctgtctttatttaggggttttgcgcgcgcggtaggcccggga ccagcggtctcggtcgttgagggtcc tgtgtattttttccaggacgtggtaaaggtgactctggatgttcagatacatgggcataa gcccgtctctggggtggaggtagcacc actgcagagcttcatgctgcggggtggtgttgtagatgatccagtcgtagcaggagcgct gggcgtggtgcctaaaaatgtctttc agtagcaagctgattgccaggggcaggcccttggtgtaagtgtttacaaagcggttaagc tgggatgggtgcatacgtggggat atgagatgcatcttggactgtatttttaggttggctatgttcccagccatatccctccgg ggattcatgttgtgcagaaccaccagca cagtgtatccggtgcacttgggaaatttgtcatgtagcttagaaggaaatgcgtggaaga acttggagacgcccttgtgacctcca agattttccatgcattcgtccataatgatggcaatgggcccacgggcggcggcctgggcg aagatatttctgggatcactaacgtc atagttgtgttccaggatgagatcgtcataggccatttttacaaagcgcgggcggagggt gccagactgcggtataatggttccat ccggcccaggggcgtagttaccctcacagatttgcatttcccacgctttgagttcagatg gggggatcatgtctacctgcggggc gatgaagaaaacggtttccggggtaggggagatcagctgggaagaaagcaggttcctgag cagctgcgacttaccgcagccg gtgggcccgtaaatcacacctattaccggctgcaactggtagttaagagagctgcagctg ccgtcatccctgagcaggggggcc acttcgttaagcatgtccctgactcgcatgttttccctgaccaaatccgccagaaggcgc tcgccgcccagcgatagcagttcttg caaggaagcaaagtttttcaacggtttgagaccgtccgccgtaggcatgcttttgagcgt ttgaccaagcagttccaggcggtccc acagctcggtcacctgctctacggcatctcgatccagcatatctcctcgtttcgcgggtt ggggcggctttcgctgtacggcagta gtcggtgctcgtccagacgggccagggtcatgtctttccacgggcgcagggtcctcgtca gcgtagtctgggtcacggtgaagg ggtgcgctccgggctgcgcgctggccagggtgcgcttgaggctggtcctgctggtgctga agcgctgccggtcttcgccctgc gcgtcggccaggtagcatttgaccatggtgtcatagtccagcccctccgcggcgtggccc ttggcgcgcagcttgcccttggag gaggcgccgcacgaggggcagtgcagacttttgagggcgtagagcttgggcgcgagaaat accgattccggggagtaggca tccgcgccgcaggccccgcagacggtctcgcattccacgagccaggtgagctctggccgt tcggggtcaaaaaccaggtttcc cccatgctttttgatgcgtttcttacctctggtttccatgagccggtgtccacgctcggt gacgaaaaggctgtccgtgtccccgtata cagacttgagaggcctgtcctcgagcggtgttccgcggtcctcctcgtatagaaactcgg accactctgagacaaaggctcgcgt ccaggccagcacgaaggaggctaagtgggaggggtagcggtcgttgtccactagggggtc cactcgctccagggtgtgaaga cacatgtcgccctcttcggcatcaaggaaggtgattggtttgtaggtgtaggccacgtga ccgggtgttcctgaaggggggctat aaaagggggtgggggcgcgttcgtcctcactctcttccgcatcgctgtctgcgagggcca gctgttggggtgagtactccctctg aaaagcgggcatgacttctgcgctaagattgtcagtttccaaaaacgaggaggatttgat attcacctggcccgcggtgatgccttt gagggtggccgcatccatctggtcagaaaagacaatctttttgttgtcaagcttggtggc aaacgacccgtagagggcgttggac agcaacttggcgatggagcgcagggtttggtttttgtcgcgatcggcgcgctccttggcc gcgatgtttagctgcacgtattcgcg cgcaacgcaccgccattcgggaaagacggtggtgcgctcgtcgggcaccaggtgcacgcg ccaaccgcggttgtgcagggt gacaaggtcaacgctggtggctacctctccgcgtaggcgctcgttggtccagcagaggcg gccgcccttgcgcgagcagaat ggcggtagggggtctagctgcgtctcgtccggggggtctgcgtccacggtaaagaccccg ggcagcaggcgcgcgtcgaag tagtctatcttgcatccttgcaagtctagcgcctgctgccatgcgcgggcggcaagcgcg cgctcgtatgggttgagtgggggac cccatggcatggggtgggtgagcgcggaggcgtacatgccgcaaatgtcgtaaacgtaga ggggctctctgagtattccaaga tatgtagggtagcatcttccaccgcggatgctggcgcgcacgtaatcgtatagttcgtgc gagggagcgaggaggtcgggacc gaggttgctacgggcgggctgctctgctcggaagactatctgcctgaagatggcatgtga gttggatgatatggttggacgctgg aagacgttgaagctggcgtctgtgagacctaccgcgtcacgcacgaaggaggcgtaggag tcgcgcagcttgttgaccagctc ggcggtgacctgcacgtctagggcgcagtagtccagggtttccttgatgatgtcatactt atcctgtcccttttttttccacagctcgc ggttgaggacaaactcttcgcggtctttccagtactcttggatcggaaacccgtcggcct ccgaacggtaagagcctagcatgta gaactggttgacggcctggtaggcgcagcatcccttttctacgggtagcgcgtatgcctg cgcggccttccggagcgaggtgtg ggtgagcgcaaaggtgtccctgaccatgactttgaggtactggtatttgaagtcagtgtc gtcgcatccgccctgctcccagagca aaaagtccgtgcgctttttggaacgcggatttggcagggcgaaggtgacatcgttgaaga gtatctttcccgcgcgaggcataaa gttgcgtgtgatgcggaagggtcccggcacctcggaacggttgttaattacctgggcggc gagcacgatctcgtcaaagccgtt gatgttgtggcccacaatgtaaagttccaagaagcgcgggatgcccttgatggaaggcaa ttttttaagttcctcgtaggtgagctc ttcaggggagctgagcccgtgctctgaaagggcccagtctgcaagatgagggttggaagc gacgaatgagctccacaggtca cgggccattagcatttgcaggtggtcgcgaaaggtcctaaactggcgacctatggccatt ttttctggggtgatgcagtagaaggt aagcgggtcttgttcccagcggtcccatccaaggttcgcggctaggtctcgcgcggcagt cactagaggctcatctccgccgaa cttcatgaccagcatgaagggcacgagctgcttcccaaaggcccccatccaagtataggt ctctacatcgtaggtgacaaagag acgctcggtgcgaggatgcgagccgatcgggaagaactggatctcccgccaccaattgga ggagtggctattgatgtggtgaa agtagaagtccctgcgacgggccgaacactcgtgctggcttttgtaaaaacgtgcgcagt actggcagcggtgcacgggctgta catcctgcacgaggttgacctgacgaccgcgcacaaggaagcagagtgggaatttgagcc cctcgcctggcgggtttggctgg tggtcttctacttcggctgcttgtccttgaccgtctggctgctcgaggggagttacggtg gatcggaccaccacgccgcgcgagc ccaaagtccagatgtccgcgcgcggcggtcggagcttgatgacaacatcgcgcagatggg agctgtccatggtctggagctcc cgcggcgtcaggtcaggcgggagctcctgcaggtttacctcgcatagacgggtcagggcg cgggctagatccaggtgatacct aatttccaggggctggttggtggcggcgtcgatggcttgcaagaggccgcatccccgcgg cgcgactacggtaccgcgcggc gggcggtgggccgcgggggtgtccttggatgatgcatctaaaagcggtgacgcgggcgag cccccggaggtagggggggc tccggacccgccgggagagggggcaggggcacgtcggcgccgcgcgcgggcaggagctgg tgctgcgcgcgtaggttgc tggcgaacgcgacgacgcggcggttgatctcctgaatctggcgcctctgcgtgaagacga cgggcccggtgagcttgaacctg aaagagagttcgacagaatcaatttcggtgtcgttgacggcggcctggcgcaaaatctcc tgcacgtctcctgagttgtcttgata ggcgatctcggccatgaactgctcgatctcttcctcctggagatctccgcgtccggctcg ctccacggtggcggcgaggtcgttg gaaatgcgggccatgagctgcgagaaggcgttgaggcctccctcgttccagacgcggctg tagaccacgcccccttcggcatc gcgggcgcgcatgaccacctgcgcgagattgagctccacgtgccgggcgaagacggcgta gtttcgcaggcgctgaaagag gtagttgagggtggtggcggtgtgttctgccacgaagaagtacataacccagcgtcgcaa cgtggattcgttgatatcccccaag gcctcaaggcgctccatggcctcgtagaagtccacggcgaagttgaaaaactgggagttg cgcgccgacacggttaactcctc ctccagaagacggatgagctcggcgacagtgtcgcgcacctcgcgctcaaaggctacagg ggcctcttcttcttcttcaatctcct cttccataagggcctccccttcttcttcttctggcggcggtgggggaggggggacacggc ggcgacgacggcgcaccgggag gcggtcgacaaagcgctcgatcatctccccgcggcgacggcgcatggtctcggtgacggc gcggccgttctcgcgggggcg cagttggaagacgccgcccgtcatgtcccggttatgggttggcggggggctgccatgcgg cagggatacggcgctaacgatg catctcaacaattgttgtgtaggtactccgccgccgagggacctgagcgagtccgcatcg accggatcggaaaacctctcgaga aaggcgtctaaccagtcacagtcgcaaggtaggctgagcaccgtggcgggcggcagcggg cggcggtcggggttgtttctgg cggaggtgctgctgatgatgtaattaaagtaggcggtcttgagacggcggatggtcgaca gaagcaccatgtccttgggtccgg cctgctgaatgcgcaggcggtcggccatgccccaggcttcgttttgacatcggcgcaggt ctttgtagtagtcttgcatgagccttt ctaccggcacttcttcttctccttcctcttgtcctgcatctcttgcatctatcgctgcgg cggcggcggagtttggccgtaggtggcg ccctcttcctcccatgcgtgtgaccccgaagcccctcatcggctgaagcagggctaggtc ggcgacaacgcgctcggctaatat ggcctgctgcacctgcgtgagggtagactggaagtcatccatgtccacaaagcggtggta tgcgcccgtgttgatggtgtaagtg cagttggccataacggaccagttaacggtctggtgacccggctgcgagagctcggtgtac ctgagacgcgagtaagccctcga gtcaaatacgtagtcgttgcaagtccgcaccaggtactggtatcccaccaaaaagtgcgg cggcggctggcggtagaggggcc agcgtagggtggccggggctccgggggcgagatcttccaacataaggcgatgatatccgt agatgtacctggacatccaggtg atgccggcggcggtggtggaggcgcgcggaaagtcgcggacgcggttccagatgttgcgc agcggcaaaaagtgctccatg gtcgggacgctctggccggtcaggcgcgcgcaatcgttgacgctctagcgtgcaaaagga gagcctgtaagcgggcactcttc cgtggtctggtggataaattcgcaagggtatcatggcggacgaccggggttcgagccccg tatccggccgtccgccgtgatcca tgcggttaccgcccgcgtgtcgaacccaggtgtgcgacgtcagacaacgggggagtgctc cttttggcttccttccaggcgcgg cggctgctgcgctagcttttttggccactggccgcgcgcagcgtaagcggttaggctgga aagcgaaagcattaagtggctcgc tccctgtagccggagggttattttccaagggttgagtcgcgggacccccggttcgagtct cggaccggccggactgcggcgaa cgggggtttgcctccccgtcatgcaagaccccgcttgcaaattcctccggaaacagggac gagccccttttttgcttttcccagat gcatccggtgctgcggcagatgcgcccccctcctcagcagcggcaagagcaagagcagcg gcagacatgcagggcaccctc ccctcctcctaccgcgtcaggaggggcgacatccgcggttgacgcggcagcagatggtga ttacgaacccccgcggcgccg ggcccggcactacctggacttggaggagggcgagggcctggcgcggctaggagcgccctc tcctgagcggcacccaaggg tgcagctgaagcgtgatacgcgtgaggcgtacgtgccgcggcagaacctgtttcgcgacc gcgagggagaggagcccgagg agatgcgggatcgaaagttccacgcagggcgcgagctgcggcatggcctgaatcgcgagc ggttgctgcgcgaggaggactt tgagcccgacgcgcgaaccgggattagtcccgcgcgcgcacacgtggcggccgccgacct ggtaaccgcatacgagcaga cggtgaaccaggagattaactttcaaaaaagctttaacaaccacgtgcgtacgcttgtgg cgcgcgaggaggtggctataggac tgatgcatctgtgggactttgtaagcgcgctggagcaaaacccaaatagcaagccgctca tggcgcagctgttccttatagtgca gcacagcagggacaacgaggcattcagggatgcgctgctaaacatagtagagcccgaggg ccgctggctgctcgatttgataa acatcctgcagagcatagtggtgcaggagcgcagcttgagcctggctgacaaggtggccg ccatcaactattccatgcttagcct gggcaagttttacgcccgcaagatataccataccccttacgttcccatagacaaggaggt aaagatcgaggggttctacatgcgc atggcgctgaaggtgcttaccttgagcgacgacctgggcgtttatcgcaacgagcgcatc cacaaggccgtgagcgtgagccg gcggcgcgagctcagcgaccgcgagctgatgcacagcctgcaaagggccctggctggcac gggcagcggcgatagagag gccgagtcctactttgacgcgggcgctgacctgcgctgggccccaagccgacgcgccctg gaggcagctggggccggacct gggctggcggtggcacccgcgcgcgctggcaacgtcggcggcgtggaggaatatgacgag gacgatgagtacgagccaga ggacggcgagtactaagcggtgatgtttctgatcagatgatgcaagacgcaacggacccg gcggtgcgggcggcgctgcaga gccagccgtccggccttaactccacggacgactggcgccaggtcatggaccgcatcatgt cgctgactgcgcgcaatcctgac gcgttccggcagcagccgcaggccaaccggctctccgcaattctggaagcggtggtcccg gcgcgcgcaaaccccacgcac gagaaggtgctggcgatcgtaaacgcgctggccgaaaacagggccatccggcccgacgag gccggcctggtctacgacgc gctgcttcagcgcgtggctcgttacaacagcggcaacgtgcagaccaacctggaccggct ggtgggggatgtgcgcgaggcc gtggcgcagcgtgagcgcgcgcagcagcagggcaacctgggctccatggttgcactaaac gccttcctgagtacacagcccg ccaacgtgccgcggggacaggaggactacaccaactttgtgagcgcactgcggctaatgg tgactgagacaccgcaaagtga ggtgtaccagtctgggccagactattttttccagaccagtagacaaggcctgcagaccgt aaacctgagccaggctttcaaaaac ttgcaggggctgtggggggtgcgggctcccacaggcgaccgcgcgaccgtgtctagcttg ctgacgcccaactcgcgcctgtt gctgctgctaatagcgcccttcacggacagtggcagcgtgtcccgggacacatacctagg tcacttgctgacactgtaccgcga ggccataggtcaggcgcatgtggacgagcatactttccaggagattacaagtgtcagccg cgcgctggggcaggaggacacg ggcagcctggaggcaaccctaaactacctgctgaccaaccggcggcagaagatcccctcg ttgcacagtttaaacagcgagga ggagcgcattttgcgctacgtgcagcagagcgtgagccttaacctgatgcgcgacggggt aacgcccagcgtggcgctggac atgaccgcgcgcaacatggaaccgggcatgtatgcctcaaaccggccgtttatcaaccgc ctaatggactacttgcatcgcgcg gccgccgtgaaccccgagtatttcaccaatgccatcttgaacccgcactggctaccgccc cctggtttctacaccgggggattcg aggtgcccgagggtaacgatggattcctctgggacgacatagacgacagcgtgttttccc cgcaaccgcagaccctgctagagt tgcaacagcgcgagcaggcagaggcggcgctgcgaaaggaaagcttccgcaggccaagca gcttgtccgatctaggcgctg cggccccgcggtcagatgctagtagcccatttccaagcttgatagggtctcttaccagca ctcgcaccacccgcccgcgcctgct gggcgaggaggagtacctaaacaactcgctgctgcagccgcagcgcgaaaaaaacctgcc tccggcatttcccaacaacggg atagagagcctagtggacaagatgagtagatggaagacgtacgcgcaggagcacagggac gtgccaggcccgcgcccgcc cacccgtcgtcaaaggcacgaccgtcagcggggtctggtgtgggaggacgatgactcggc agacgacagcagcgtcctggat ttgggagggagtggcaacccgtttgcgcaccttcgccccaggctggggagaatgttttaa aaaaaaaaaagcatgatgcaaaat aaaaaactcaccaaggccatggcaccgagcgttggttttcttgtattccccttagtatgc ggcgcgcggcgatgtatgaggaaggt cctcctccctcctacgagagtgtggtgagcgcggcgccagtggcggcggcgctgggttct cccttcgatgctcccctggacccg ccgtttgtgcctccgcggtacctgcggcctaccggggggagaaacagcatccgttactct gagttggcacccctattcgacacca cccgtgtgtacctggtggacaacaagtcaacggatgtggcatccctgaactaccagaacg accacagcaactttctgaccacgg tcattcaaaacaatgactacagcccgggggaggcaagcacacagaccatcaatcttgacg accggtcgcactggggcggcga cctgaaaaccatcctgcataccaacatgccaaatgtgaacgagttcatgtttaccaataa gtttaaggcgcgggtgatggtgtcgc gcttgcctactaaggacaatcaggtggagctgaaatacgagtgggtggagttcacgctgc ccgagggcaactactccgagacc atgaccatagaccttatgaacaacgcgatcgtggagcactacttgaaagtgggcagacag aacggggttctggaaagcgacat cggggtaaagtttgacacccgcaacttcagactggggtttgaccccgtcactggtcttgt catgcctggggtatatacaaacgaag ccttccatccagacatcattttgctgccaggatgcggggtggacttcacccacagccgcc tgagcaacttgttgggcatccgcaa gcggcaacccttccaggagggctttaggatcacctacgatgatctggagggtggtaacat tcccgcactgttggatgtggacgcc taccaggcgagcttgaaagatgacaccgaacagggcgggggtggcgcaggcggcagcaac agcagtggcagcggcgcgg aagagaactccaacgcggcagccgcggcaatgcagccggtggaggacatgaacgatcatg ccattcgcggcgacacctttgc cacacgggctgaggagaagcgcgctgaggccgaagcagcggccgaagctgccgcccccgc tgcgcaacccgaggtcgag aagcctcagaagaaaccggtgatcaaacccctgacagaggacagcaagaaacgcagttac aacctaataagcaatgacagca ccttcacccagtaccgcagctggtaccttgcatacaactacggcgaccctcagaccggaa tccgctcatggaccctgctttgcac tcctgacgtaacctgcggctcggagcaggtctactggtcgttgccagacatgatgcaaga ccccgtgaccttccgctccacgcg ccagatcagcaactttccggtggtgggcgccgagctgttgcccgtgcactccaagagctt ctacaacgaccaggccgtctactc ccaactcatccgccagtttacctctctgacccacgtgttcaatcgctttcccgagaacca gattttggcgcgcccgccagccccca ccatcaccaccgtcagtgaaaacgttcctgctctcacagatcacgggacgctaccgctgc gcaacagcatcggaggagtccag cgagtgaccattactgacgccagacgccgcacctgcccctacgtttacaaggccctgggc atagtctcgccgcgcgtcctatcg agccgcactttttgagcaagcatgtccatccttatatcgcccagcaataacacaggctgg ggcctgcgcttcccaagcaagatgtt tggcggggccaagaagcgctccgaccaacacccagtgcgcgtgcgcgggcactaccgcgc gccctggggcgcgcacaaac gcggccgcactgggcgcaccaccgtcgatgacgccatcgacgcggtggtggaggaggcgc gcaactacacgcccacgccg ccaccagtgtccacagtggacgcggccattcagaccgtggtgcgcggagcccggcgctat gctaaaatgaagagacggcgg aggcgcgtagcacgtcgccaccgccgccgacccggcactgccgcccaacgcgcggcggcg gccctgcttaaccgcgcacg tcgcaccggccgacgggcggccatgcgggccgctcgaaggctggccgcgggtattgtcac tgtgccccccaggtccaggcg acgagcggccgccgcagcagccgcggccattagtgctatgactcagggtcgcaggggcaa cgtgtattgggtgcgcgactcg gttagcggcctgcgcgtgcccgtgcgcacccgccccccgcgcaactagattgcaagaaaa aactacttagactcgtactgttgta tgtatccagcggcggcggcgcgcaacgaagctatgtccaagcgcaaaatcaaagaagaga tgctccaggtcatcgcgccgga gatctatggccccccgaagaaggaagagcaggattacaagccccgaaagctaaagcgggt caaaaagaaaaagaaagatga tgatgatgaacttgacgacgaggtggaactgctgcacgctaccgcgcccaggcgacgggt acagtggaaaggtcgacgcgta aaacgtgttttgcgacccggcaccaccgtagtctttacgcccggtgagcgctccacccgc acctacaagcgcgtgtatgatgag gtgtacggcgacgaggacctgcttgagcaggccaacgagcgcctcggggagtttgcctac ggaaagcggcataaggacatgc tggcgttgccgctggacgagggcaacccaacacctagcctaaagcccgtaacactgcagc aggtgctgcccgcgcttgcacc gtccgaagaaaagcgcggcctaaagcgcgagtctggtgacttggcacccaccgtgcagct gatggtacccaagcgccagcga ctggaagatgtcttggaaaaaatgaccgtggaacctgggctggagcccgaggtccgcgtg cggccaatcaagcaggtggcgc cgggactgggcgtgcagaccgtggacgttcagatacccactaccagtagcaccagtattg ccaccgccacagagggcatgga gacacaaacgtccccggttgcctcagcggtggcggatgccgcggtgcaggcggtcgctgc ggccgcgtccaagacctctacg gaggtgcaaacggacccgtggatgtttcgcgtttcagccccccggcgcccgcgccgttcg aggaagtacggcgccgccagcg cgctactgcccgaatatgccctacatccttccattgcgcctacccccggctatcgtggct acacctaccgccccagaagacgagc aactacccgacgccgaaccaccactggaacccgccgccgccgtcgccgtcgccagcccgt gctggccccgatttccgtgcgc agggtggctcgcgaaggaggcaggaccctggtgctgccaacagcgcgctaccaccccagc atcgtttaaaagccggtctttgt ggttcttgcagatatggccctcacctgccgcctccgtttcccggtgccgggattccgagg aagaatgcaccgtaggaggggcat ggccggccacggcctgacgggcggcatgcgtcgtgcgcaccaccggcggcggcgcgcgtc gcaccgtcgcatgcgcggc ggtatcctgcccctccttattccactgatcgccgcggcgattggcgccgtgcccggaatt gcatccgtggccttgcaggcgcaga gacactgattaaaaacaagttgcatgtggaaaaatcaaaataaaaagtctggactctcac gctcgcttggtcctgtaactattttgta gaatggaagacatcaactttgcgtctctggccccgcgacacggctcgcgcccgttcatgg gaaactggcaagatatcggcacca gcaatatgagcggtggcgccttcagctggggctcgctgtggagcggcattaaaaatttcg gttccaccgttaagaactatggcag caaggcctggaacagcagcacaggccagatgctgagggataagttgaaagagcaaaattt ccaacaaaaggtggtagatggc ctggcctctggcattagcggggtggtggacctggccaaccaggcagtgcaaaataagatt aacagtaagcttgatccccgccct cccgtagaggagcctccaccggccgtggagacagtgtctccagaggggcgtggcgaaaag cgtccgcgccccgacaggga agaaactctggtgacgcaaatagacgagcctccctcgtacgaggaggcactaaagcaagg cctgcccaccacccgtcccatc gcgcccatggctaccggagtgctgggccagcacacacccgtaacgctggacctgcctccc cccgccgacacccagcagaaa cctgtgctgccaggcccgaccgccgttgttgtaacccgtcctagccgcgcgtccctgcgc cgcgccgccagcggtccgcgatc gttgcggcccgtagccagtggcaactggcaaagcacactgaacagcatcgtgggtctggg ggtgcaatccctgaagcgccga cgatgcttctgatagctaacgtgtcgtatgtgtgtcatgtatgcgtccatgtcgccgcca gaggagctgctgagccgccgcgcgc ccgctttccaagatggctaccccttcgatgatgccgcagtggtcttacatgcacatctcg ggccaggacgcctcggagtacctga gccccgggctggtgcagtttgcccgcgccaccgagacgtacttcagcctgaataacaagt ttagaaaccccacggtggcgccta cgcacgacgtgaccacagaccggtcccagcgtttgacgctgcggttcatccctgtggacc gtgaggatactgcgtactcgtaca aggcgcggttcaccctagctgtgggtgataaccgtgtgctggacatggcttccacgtact ttgacatccgcggcgtgctggacag gggccctacttttaagccctactctggcactgcctacaacgccctggctcccaagggtgc cccaaatccttgcgaatgggatgaa gctgctactgctcttgaaataaacctagaagaagaggacgatgacaacgaagacgaagta gacgagcaagctgagcagcaaa aaactcacgtatttgggcaggcgccttattctggtataaatattacaaaggagggtattc aaataggtgtcgaaggtcaaacaccta aatatgccgataaaacatttcaacctgaacctcaaataggagaatctcagtggtacgaaa cagaaattaatcatgcagctgggag agtcctaaaaaagactaccccaatgaaaccatgttacggttcatatgcaaaacccacaaa tgaaaatggagggcaaggcattctt gtaaagcaacaaaatggaaagctagaaagtcaagtggaaatgcaatttttctcaactact gaggcagccgcaggcaatggtgata acttgactcctaaagtggtattgtacagtgaagatgtagatatagaaaccccagacactc atatttcttacatgcccactattaagga aggtaactcacgagaactaatgggccaacaatctatgcccaacaggcctaattacattgc ttttagggacaattttattggtctaatg tattacaacagcacgggtaatatgggtgttctggcgggccaagcatcgcagttgaatgct gttgtagatttgcaagacagaaacac agagctttcataccagcttttgcttgattccattggtgatagaaccaggtacttttctat gtggaatcaggctgttgacagctatgatcc agatgttagaattattgaaaatcatggaactgaagatgaacttccaaattactgctttcc actgggaggtgtgattaatacagagact cttaccaaggtaaaacctaaaacaggtcaggaaaatggatgggaaaaagatgctacagaa ttttcagataaaaatgaaataagag ttggaaataattttgccatggaaatcaatctaaatgccaacctgtggagaaatttcctgt actccaacatagcgctgtatttgcccgac aagctaaagtacagtccttccaacgtaaaaatttctgataacccaaacacctacgactac atgaacaagcgagtggtggctcccg ggctagtggactgctacattaaccttggagcacgctggtcccttgactatatggacaacg tcaacccatttaaccaccaccgcaat gctggcctgcgctaccgctcaatgttgctgggcaatggtcgctatgtgcccttccacatc caggtgcctcagaagttctttgccatt aaaaacctccttctcctgccgggctcatacacctacgagtggaacttcaggaaggatgtt aacatggttctgcagagctccctagg aaatgacctaagggttgacggagccagcattaagtttgatagcatttgcctttacgccac cttcttccccatggcccacaacaccgc ctccacgcttgaggccatgcttagaaacgacaccaacgaccagtcctttaacgactatct ctccgccgccaacatgctctacccta tacccgccaacgctaccaacgtgcccatatccatcccctcccgcaactgggcggctttcc gcggctgggccttcacgcgccttaa gactaaggaaaccccatcactgggctcgggctacgacccttattacacctactctggctc tataccctacctagatggaacctttta cctcaaccacacctttaagaaggtggccattacctttgactcttctgtcagctggcctgg caatgaccgcctgcttacccccaacga gtttgaaattaagcgctcagttgacggggagggttacaacgttgcccagtgtaacatgac caaagactggttcctggtacaaatgc tagctaactataacattggctaccagggcttctatatcccagagagctacaaggaccgca tgtactccttctttagaaacttccagcc catgagccgtcaggtggtggatgatactaaatacaaggactaccaacaggtgggcatcct acaccaacacaacaactctggattt gttggctaccttgcccccaccatgcgcgaaggacaggcctaccctgctaacttcccctat ccgcttataggcaagaccgcagttg acagcattacccagaaaaagtttctttgcgatcgcaccctttggcgcatcccattctcca gtaactttatgtccatgggcgcactcac agacctgggccaaaaccttctctacgccaactccgcccacgcgctagacatgacttttga ggtggatcccatggacgagcccac ccttctttatgttttgtttgaagtctttgacgtggtccgtgtgcaccagccgcaccgcgg cgtcatcgaaaccgtgtacctgcgcacg cccttctcggccggcaacgccacaacataaagaagcaagcaacatcaacaacagctgccg ccatgggctccagtgagcagga actgaaagccattgtcaaagatcttggttgtgggccatattttttgggcacctatgacaa gcgctttccaggctttgtttctccacaca agctcgcctgcgccatagtcaatacggccggtcgcgagactgggggcgtacactggatgg cctttgcctggaacccgcactca aaaacatgctacctctttgagccctttggcttttctgaccagcgactcaagcaggtttac cagtttgagtacgagtcactcctgcgcc gtagcgccattgcttcttcccccgaccgctgtataacgctggaaaagtccacccaaagcg tacaggggcccaactcggccgcct gtggactattctgctgcatgtttctccacgcctttgccaactggccccaaactcccatgg atcacaaccccaccatgaaccttattac cggggtacccaactccatgctcaacagtccccaggtacagcccaccctgcgtcgcaacca ggaacagctctacagcttcctgg agcgccactcgccctacttccgcagccacagtgcgcagattaggagcgccacttcttttt gtcacttgaaaaacatgtaaaaataat gtactagagacactttcaataaaggcaaatgcttttatttgtacactctcgggtgattat ttacccccacccttgccgtctgcgccgttt aaaaatcaaaggggttctgccgcgcatcgctatgcgccactggcagggacacgttgcgat actggtgtttagtgctccacttaaa ctcaggcacaaccatccgcggcagctcggtgaagttttcactccacaggctgcgcaccat caccaacgcgtttagcaggtcggg cgccgatatcttgaagtcgcagttggggcctccgccctgcgcgcgcgagttgcgatacac agggttgcagcactggaacactat cagcgccgggtggtgcacgctggccagcacgctcttgtcggagatcagatccgcgtccag gtcctccgcgttgctcagggcga acggagtcaactttggtagctgccttcccaaaaagggcgcgtgcccaggctttgagttgc actcgcaccgtagtggcatcaaaag gtgaccgtgcccggtctgggcgttaggatacagcgcctgcataaaagccttgatctgctt aaaagccacctgagcctttgcgcctt cagagaagaacatgccgcaagacttgccggaaaactgattggccggacaggccgcgtcgt gcacgcagcaccttgcgtcggt gttggagatctgcaccacatttcggccccaccggttcttcacgatcttggccttgctaga ctgctccttcagcgcgcgctgcccgttt tcgctcgtcacatccatttcaatcacgtgctccttatttatcataatgcttccgtgtaga cacttaagctcgccttcgatctcagcgcag cggtgcagccacaacgcgcagcccgtgggctcgtgatgcttgtaggtcacctctgcaaac gactgcaggtacgcctgcaggaa tcgccccatcatcgtcacaaaggtcttgttgctggtgaaggtcagctgcaacccgcggtg ctcctcgttcagccaggtcttgcata cggccgccagagcttccacttggtcaggcagtagtttgaagttcgcctttagatcgttat ccacgtggtacttgtccatcagcgcgc gcgcagcctccatgcccttctcccacgcagacacgatcggcacactcagcgggttcatca ccgtaatttcactttccgcttcgctg ggctcttcctcttcctcttgcgtccgcataccacgcgccactgggtcgtcttcattcagc cgccgcactgtgcgcttacctcctttgc catgcttgattagcaccggtgggttgctgaaacccaccatttgtagcgccacatcttctc tttcttcctcgctgtccacgattacctctg gtgatggcgggcgctcgggcttgggagaagggcgcttctttttcttcttgggcgcaatgg ccaaatccgccgccgaggtcgatg gccgcgggctgggtgtgcgcggcaccagcgcgtcttgtgatgagtcttcctcgtcctcgg actcgatacgccgcctcatccgctt ttttgggggcgcccggggaggcggcggcgacggggacggggacgacacgtcctccatggt tgggggacgtcgcgccgcac cgcgtccgcgctcgggggtggtttcgcgctgctcctcttcccgactggccatttccttct cctataggcagaaaaagatcatggag tcagtcgagaagaaggacagcctaaccgccccctctgagttcgccaccaccgcctccacc gatgccgccaacgcgcctacca ccttccccgtcgaggcacccccgcttgaggaggaggaagtgattatcgagcaggacccag gttttgtaagcgaagacgacgag gaccgctcagtaccaacagaggataaaaagcaagaccaggacaacgcagaggcaaacgag gaacaagtcgggcgggggg acgaaaggcatggcgactacctagatgtgggagacgacgtgctgttgaagcatctgcagc gccagtgcgccattatctgcgac gcgttgcaagagcgcagcgatgtgcccctcgccatagcggatgtcagccttgcctacgaa cgccacctattctcaccgcgcgta ccccccaaacgccaagaaaacggcacatgcgagcccaacccgcgcctcaacttctacccc gtatttgccgtgccagaggtgctt gccacctatcacatctttttccaaaactgcaagatacccctatcctgccgtgccaaccgc agccgagcggacaagcagctggcct tgcggcagggcgctgtcatacctgatatcgcctcgctcaacgaagtgccaaaaatctttg agggtcttggacgcgacgagaagc gcgcggcaaacgctctgcaacaggaaaacagcgaaaatgaaagtcactctggagtgttgg tggaactcgagggtgacaacgc gcgcctagccgtactaaaacgcagcatcgaggtcacccactttgcctacccggcacttaa cctaccccccaaggtcatgagcac agtcatgagtgagctgatcgtgcgccgtgcgcagcccctggagagggatgcaaatttgca agaacaaacagaggagggccta cccgcagttggcgacgagcagctagcgcgctggcttcaaacgcgcgagcctgccgacttg gaggagcgacgcaaactaatga tggccgcagtgctcgttaccgtggagcttgagtgcatgcagcggttctttgctgacccgg agatgcagcgcaagctagaggaaa cattgcactacacctttcgacagggctacgtacgccaggcctgcaagatctccaacgtgg agctctgcaacctggtctcctacctt ggaattttgcacgaaaaccgccttgggcaaaacgtgcttcattccacgctcaagggcgag gcgcgccgcgactacgtccgcga ctgcgtttacttatttctatgctacacctggcagacggccatgggcgtttggcagcagtg cttggaggagtgcaacctcaaggagc tgcagaaactgctaaagcaaaacttgaaggacctatggacggccttcaacgagcgctccg tggccgcgcacctggcggacatc attttccccgaacgcctgcttaaaaccctgcaacagggtctgccagacttcaccagtcaa agcatgttgcagaactttaggaacttt atcctagagcgctcaggaatcttgcccgccacctgctgtgcacttcctagcgactttgtg cccattaagtaccgcgaatgccctcc gccgctttggggccactgctaccttctgcagctagccaactaccttgcctaccactctga cataatggaagacgtgagcggtgac ggtctactggagtgtcactgtcgctgcaacctatgcaccccgcaccgctccctggtttgc aattcgcagctgcttaacgaaagtca aattatcggtacctttgagctgcagggtccctcgcctgacgaaaagtccgcggctccggg gttgaaactcactccggggctgtgg acgtcggcttaccttcgcaaatttgtacctgaggactaccacgcccacgagattaggttc tacgaagaccaatcccgcccgccta atgcggagcttaccgcctgcgtcattacccagggccacattcttggccaattgcaagcca tcaacaaagcccgccaagagtttct gctacgaaagggacggggggtttacttggacccccagtccggcgaggagctcaacccaat ccccccgccgccgcagccctat cagcagcagccgcgggcccttgcttcccaggatggcacccaaaaagaagctgcagctgcc gccgccacccacggacgagg aggaatactgggacagtcaggcagaggaggttttggacgaggaggaggaggacatgatgg aagactgggagagcctagacg aggaagcttccgaggtcgaagaggtgtcagacgaaacaccgtcaccctcggtcgcattcc cctcgccggcgccccagaaatc ggcaaccggttccagcatggctacaacctccgctcctcaggcgccgccggcactgcccgt tcgccgacccaaccgtagatgg gacaccactggaaccagggccggtaagtccaagcagccgccgccgttagcccaagagcaa caacagcgccaaggctaccg ctcatggcgcgggcacaagaacgccatagttgcttgcttgcaagactgtgggggcaacat ctccttcgcccgccgctttcttctct accatcacggcgtggccttcccccgtaacatcctgcattactaccgtcatctctacagcc catactgcaccggcggcagcggcag caacagcagcggccacacagaagcaaaggcgaccggatagcaagactctgacaaagccca agaaatccacagcggcggca gcagcaggaggaggagcgctgcgtctggcgcccaacgaacccgtatcgacccgcgagctt agaaacaggatttttcccactct gtatgctatatttcaacagagcaggggccaagaacaagagctgaaaataaaaaacaggtc tctgcgatccctcacccgcagctg cctgtatcacaaaagcgaagatcagcttcggcgcacgctggaagacgcggaggctctctt cagtaaatactgcgcgctgactctt aaggactagtttcgcgccctttctcaaatttaagcgcgaaaactacgtcatctccagcgg ccacacccggcgccagcacctgttgt cagcgccattatgagcaaggaaattcccacgccctacatgtggagttaccagccacaaat gggacttgcggctggagctgccca agactactcaacccgaataaactacatgagcgcgggaccccacatgatatcccgggtcaa cggaatacgcgcccaccgaaac cgaattctcctggaacaggcggctattaccaccacacctcgtaataaccttaatccccgt agttggcccgctgccctggtgtacca ggaaagtcccgctcccaccactgtggtacttcccagagacgcccaggccgaagttcagat gactaactcaggggcgcagcttg cgggcggctttcgtcacagggtgcggtcgcccgggcagggtataactcacctgacaatca gagggcgaggtattcagctcaac gacgagtcggtgagctcctcgcttggtctccgtccggacgggacatttcagatcggcggc gccggccgctcttcattcacgcctc gtcaggcaatcctaactctgcagacctcgtcctctgagccgcgctctggaggcattggaa ctctgcaatttattgaggagtttgtgc catcggtctactttaaccccttctcgggacctcccggccactatccggatcaatttattc ctaactttgacgcggtaaaggactcggc ggacggctacgactgaatgttaagtggagaggcagagcaactgcgcctgaaacacctggt ccactgtcgccgccacaagtgct ttgcccgcgactccggtgagttttgctactttgaattgcccgaggatcatatcgagggcc cggcgcacggcgtccggcttaccgc ccagggagagcttgcccgtagcctgattcgggagtttacccagcgccccctgctagttga gcgggacaggggaccctgtgttct cactgtgatttgcaactgtcctaaccctggattacatcaagatcctctagttaatgtcag gtcgcctaagtcgattaactagagtaccc ggggatcttattccctttaactaataaaaaaaaataataaagcatcacttacttaaaatc agttagcaaatttctgtccagtttattcagc agcacctccttgccctcctcccagctctggtattgcagcttcctcctggctgcaaacttt ctccacaatctaaatggaatgtcagtttc ctcctgttcctgtccatccgcacccactatcttcatgttgttgcagatgaagcgcgcaag accgtctgaagataccttcaaccccgt gtatccatatgacacggaaaccggtcctccaactgtgccttttcttactcctccctttgt atcccccaatgggtttcaagagagtcccc ctggggtactctctttgcgcctatccgaacctctagttacctccaatggcatgcttgcgc tcaaaatgggcaacggcctctctctgg acgaggccggcaaccttacctcccaaaatgtaaccactgtgagcccacctctcaaaaaaa ccaagtcaaacataaacctggaaa tatctgcacccctcacagttacctcagaagccctaactgtggctgccgccgcacctctaa tggtcgcgggcaacacactcaccat gcaatcacaggccccgctaaccgtgcacgactccaaacttagcattgccacccaaggacc cctcacagtgtcagaaggaaagc tagccctgcaaacatcaggccccctcaccaccaccgatagcagtacccttactatcactg cctcaccccctctaactactgccact ggtagcttgggcattgacttgaaagagcccatttatacacaaaatggaaaactaggacta aagtacggggctcctttgcatgtaac agacgacctaaacactttgaccgtagcaactggtccaggtgtgactattaataatacttc cttgcaaactaaagttactggagccttg ggttttgattcacaaggcaatatgcaacttaatgtagcaggaggactaaggattgattct caaaacagacgccttatacttgatgtta gttatccgtttgatgctcaaaaccaactaaatctaagactaggacagggccctcttttta taaactcagcccacaacttggatattaac tacaacaaaggcctttacttgtttacagcttcaaacaattccaaaaagcttgaggttaac ctaagcactgccaaggggttgatgtttg acgctacagccatagccattaatgcaggagatgggcttgaatttggttcacctaatgcac caaacacaaatcccctcaaaacaaa aattggccatggcctagaatttgattcaaacaaggctatggttcctaaactaggaactgg ccttagttttgacagcacaggtgccatt acagtaggaaacaaaaataatgataagctaactttgtggaccacaccagctccatctcct aactgtagactaaatgcagagaaag atgctaaactcactttggtcttaacaaaatgtggcagtcaaatacttgctacagtttcag ttttggctgttaaaggcagtttggctccaa tatctggaacagttcaaagtgctcatcttattataagatttgacgaaaatggagtgctac taaacaattccttcctggacccagaatat tggaactttagaaatggagatcttactgaaggcacagcctatacaaacgctgttggattt atgcctaacctatcagcttatccaaaat ctcacggtaaaactgccaaaagtaacattgtcagtcaagtttacttaaacggagacaaaa ctaaacctgtaacactaaccattacac taaacggtacacaggaaacaggagacacaactccaagtgcatactctatgtcattttcat gggactggtctggccacaactacatt aatgaaatatttgccacatcctcttacactttttcatacattgcccaagaataaagaatc gtttgtgttatgtttcaacgtgtttatttttcaa ttgcagaaaatttcaagtcatttttcattcagtagtatagccccaccaccacatagctta tacagatcaccgtaccttaatcaaactca cagaaccctagtattcaacctgccacctccctcccaacacacagagtacacagtcctttc tccccggctggccttaaaaagcatca tatcatgggtaacagacatattcttaggtgttatattccacacggtttcctgtcgagcca aacgctcatcagtgatattaataaactcc ccgggcagctcacttaagttcatgtcgctgtccagctgctgagccacaggctgctgtcca acttgcggttgcttaacgggcggcg aaggagaagtccacgcctacatgggggtagagtcataatcgtgcatcaggatagggcggt ggtgctgcagcagcgcgcgaat aaactgctgccgccgccgctccgtcctgcaggaatacaacatggcagtggtctcctcagc gatgattcgcaccgcccgcagcat aaggcgccttgtcctccgggcacagcagcgcaccctgatctcacttaaatcagcacagta actgcagcacagcaccacaatatt gttcaaaatcccacagtgcaaggcgctgtatccaaagctcatggcggggaccacagaacc cacgtggccatcataccacaagc gcaggtagattaagtggcgacccctcataaacacgctggacataaacattacctcttttg gcatgttgtaattcaccacctcccggt accatataaacctctgattaaacatggcgccatccaccaccatcctaaaccagctggcca aaacctgcccgccggctatacactg cagggaaccgggactggaacaatgacagtggagagcccaggactcgtaaccatggatcat catgctcgtcatgatatcaatgtt ggcacaacacaggcacacgtgcatacacttcctcaggattacaagctcctcccgcgttag aaccatatcccagggaacaaccca ttcctgaatcagcgtaaatcccacactgcagggaagacctcgcacgtaactcacgttgtg cattgtcaaagtgttacattcgggca gcagcggatgatcctccagtatggtagcgcgggtttctgtctcaaaaggaggtagacgat ccctactgtacggagtgcgccgag acaaccgagatcgtgttggtcgtagtgtcatgccaaatggaacgccggacgtagtcatat ttcctgaagcaaaaccaggtgcgg gcgtgacaaacagatctgcgtctccggtctcgccgcttagatcgctctgtgtagtagttg tagtatatccactctctcaaagcatcca ggcgccccctggcttcgggttctatgtaaactccttcatgcgccgctgccctgataacat ccaccaccgcagaataagccacacc cagccaacctacacattcgttctgcgagtcacacacgggaggagcgggaagagctggaag aaccatgtttttttttttattccaaaa gattatccaaaacctcaaaatgaagatctattaagtgaacgcgctcccctccggtggcgt ggtcaaactctacagccaaagaaca gataatggcatttgtaagatgttgcacaatggcttccaaaaggcaaacggccctcacgtc caagtggacgtaaaggctaaaccct tcagggtgaatctcctctataaacattccagcaccttcaaccatgcccaaataattctca tctcgccaccttctcaatatatctctaagc aaatcccgaatattaagtccggccattgtaaaaatctgctccagagcgccctccaccttc agcctcaagcagcgaatcatgattgc aaaaattcaggttcctcacagacctgtataagattcaaaagcggaacattaacaaaaata ccgcgatcccgtaggtcccttcgcag ggccagctgaacataatcgtgcaggtctgcacggaccagcgcggccacttccccgccagg aaccatgacaaaagaacccaca ctgattatgacacgcatactcggagctatgctaaccagcgtagccccgatgtaagcttgt tgcatgggcggcgatataaaatgcaa ggtgctgctcaaaaaatcaggcaaagcctcgcgcaaaaaagaaagcacatcgtagtcatg ctcatgcagataaaggcaggtaa gctccggaaccaccacagaaaaagacaccatttttctctcaaacatgtctgcgggtttct gcataaacacaaaataaaataacaaa aaaacatttaaacattagaagcctgtcttacaacaggaaaaacaacccttataagcataa gacggactacggccatgccggcgtg accgtaaaaaaactggtcaccgtgattaaaaagcaccaccgacagctcctcggtcatgtc cggagtcataatgtaagactcggta aacacatcaggttgattcacatcggtcagtgctaaaaagcgaccgaaatagcccggggga atacatacccgcaggcgtagaga caacattacagcccccataggaggtataacaaaattaataggagagaaaaacacataaac acctgaaaaaccctcctgcctagg caaaatagcaccctcccgctccagaacaacatacagcgcttccacagcggcagccataac agtcagccttaccagtaaaaaag aaaacctattaaaaaaacaccactcgacacggcaccagctcaatcagtcacagtgtaaaa aagggccaagtgcagagcgagta tatataggactaaaaaatgacgtaacggttaaagtccacaaaaaacacccagaaaaccgc acgcgaacctacgcccagaaacg aaagccaaaaaacccacaacttcctcaaatcgtcacttccgttttcccacgttacgtcac ttcccattttaagaaaactacaattccca acacatacaagttactccgccctaaaacctacgtcacccgccccgttcccacgccccgcg ccacgtcacaaactccaccccctc attatcatattggcttcaatccaaaataaggtatattattgatgatgttaattaacatgc atggatcctacgtctcgaccgatgcccttga gagccttcaacccagtcagctccttccggtgggcgcggggcatgactatcgtcgccgcac ttatgactgtcttctttatcatgcaac tcgtaggacaggtgccggcagcgctctgggtcattttcggcgaggaccgctttcgctgga gcgcgacgatgatcggcctgtcgc ttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcactggtcccgccacca aacgtttcggcgagaagcaggccatt atcgccggcatggcggccgacgcgctgggctacgtcttgctggcgttcgcgacgcgaggc tggatggccttccccattatgatt cttctcgcttccggcggcatcgggatgcccgcgttgcaggccatgctgtccaggcaggta gatgacgaccatcagggacagctt caaggatcgctcgcggctcttaccagcctaacttcgatcattggaccgctgatcgtcacg gcgatttatgccgcctcggcgagca catggaacgggttggcatggattgtaggcgccgccctataccttgtctgcctccccgcgt tgcgtcgcggtgcatggagccggg ccacctcgacctgaatggaagccggcggcacctcgctaacggattcaccactccaagaat tggagccaatcaattcttgcggag aactgtgaatgcgcaaaccaacccttggcagaacatatccatcgcgtccgccatctccag cagccgcacgcggcgcatctcgg gcagcgttgggtcctggccacgggtgcgcatgatcgtgctcctgtcgttgaggacccggc taggctggcggggttgccttactg gttagcagaatgaatcaccgatacgcgagcgaacgtgaagcgactgctgctgcaaaacgt ctgcgacctgagcaacaacatga atggtcttcggtttccgtgtttcgtaaagtctggaaacgcggaagtcagcgccctgcacc attatgttccggatctgcatcgcagga tgctgctggctaccctgtggaacacctacatctgtattaacgaagcgctggcattgaccc tgagtgatttttctctggtcccgccgca tccataccgccagttgtttaccctcacaacgttccagtaaccgggcatgttcatcatcag taacccgtatcgtgagcatcctctctcg tttcatcggtatcattacccccatgaacagaaatcccccttacacggaggcatcagtgac caaacaggaaaaaaccgcccttaac atggcccgctttatcagaagccagacattaacgcttctggagaaactcaacgagctggac gcggatgaacaggcagacatctgt gaatcgcttcacgaccacgctgatgagctttaccgcagctgcctcgcgcgtttcggtgat gacggtgaaaacctctgacacatgc agctcccggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtc agggcgcgtcagcgggtgttgg cgggtgtcggggcgcagccatgacccagtcacgtagcgatagcggagtgtatactggctt aactatgcggcatcagagcagatt gtactgagagtgcaccatatgcggtgtgaaataccgcacagatgcgtaaggagaaaatac cgcatcaggcgctcttccgcttcct cgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctcactcaa aggcggtaatacggttatccacag aatcaggggataacgcaggaaagaacatgtgagcaaaaggccagcaaaaggccaggaacc gtaaaaaggccgcgttgctgg cgtttttccataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcaga ggtggcgaaacccgacaggactat aaagataccaggcgtttccccctggaagctccctcgtgcgctctcctgttccgaccctgc cgcttaccggatacctgtccgcctttc tcccttcgggaagcgtggcgctttctcatagctcacgctgtaggtatctcagttcggtgt aggtcgttcgctccaagctgggctgtgt gcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatcgtcttgagtc caacccggtaagacacgacttatcg ccactggcagcagccactggtaacaggattagcagagcgaggtatgtaggcggtgctaca gagttcttgaagtggtggcctaac tacggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttc ggaaaaagagttggtagctcttgatcc ggcaaacaaaccaccgctggtagcggtggtttttttgtttgcaagcagcagattacgcgc agaaaaaaaggatctcaagaagatc ctttgatcttttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattt tggtcatgagattatcaaaaaggatcttc acctagatccttttaaattaaaaatgaagttttaaatcaatctaaagtatatatgagtaa acttggtctgacagttaccaatgcttaatca gtgaggcacctatctcagcgatctgtctatttcgttcatccatagttgcctgactccccg tcgtgtagataactacgatacgggagg gcttaccatctggccccagtgctgcaatgataccgcgagacccacgctcaccggctccag atttatcagcaataaaccagccag ccggaagggccgagcgcagaagtggtcctgcaactttatccgcctccatccagtctatta attgttgccgggaagctagagtaag tagttcgccagttaatagtttgcgcaacgttgttgccattgctgcaggcatcgtggtgtc acgctcgtcgtttggtatggcttcattca gctccggttcccaacgatcaaggcgagttacatgatcccccatgttgtgcaaaaaagcgg ttagctccttcggtcctccgatcgtt gtcagaagtaagttggccgcagtgttatcactcatggttatggcagcactgcataattct cttactgtcatgccatccgtaagatgctt ttctgtgactggtgagtactcaaccaagtcattctgagaatagtgtatgcggcgaccgag ttgctcttgcccggcgtcaacacggg ataataccgcgccacatagcagaactttaaaagtgctcatcattggaaaacgttcttcgg ggcgaaaactctcaaggatcttaccg ctgttgagatccagttcgatgtaacccactcgtgcacccaactgatcttcagcatctttt actttcaccagcgtttctgggtgagcaaa aacaggaaggcaaaatgccgcaaaaaagggaataagggcgacacggaaatgttgaatact catactcttcctttttcaatattatt gaagcatttatcagggttattgtctcatgagcggatacatatttgaatgtatttagaaaa ataaacaaataggggttccgcgcacattt ccccgaaaagtgccacctgtctagctacgaattcttcgacagcttcgaaactagaatcga tttcgaaactagcttaagggtgggaa agaatatataaggtgggggtcttatgtagttttgtatctgttttgcagcagccgccgccg ccatgagcaccaactcgtttgatggaa gc pAD _Track8509 Adenovirus Packaging or Helper (SEQ ID NO: 8) ttaattaannntcccttccagctctctgccccttttggattgaagccaatatgataatga gggggtggagtttgtgacgtggcgcgg ggcgtgggaacggggcgggtgacgtagtagtgtggcggaagtgtgatgttgcaagtgtgg cggaacacatgtaagcgacgga tgtggcaaaagtgacgtttttggtgtgcgccggtgtacacaggaagtgacaattttcgcg cggttttaggcggatgttgtagtaaatt tgggcgtaaccgagtaagatttggccattttcgcgggaaaactgaataagaggaagtgaa atctgaataattttgtgttactcatag cgcgtaannncgcgttaagatacattgatgagtttggacaaaccacaactagaatgcagt gaaaaaaatgctttatttgtgaaattt gtgatgctattgctttatttgtaaccattataagctgcaataaacaagttaacaacaaca attgcattcattttatgtttcaggttcaggg ggaggtgtgggaggttttttaaagcaagtaaaacctctacaaatgtggtatggctgatta tgatcagttatctagatccggtggatct gagtccggacttgtacagctcgtccatgccgagagtgatcccggcggcggtcacgaactc cagcaggaccatgtgatcgcgctt ctcgttggggtctttgctcagggcggactgggtgctcaggtagtggttgtcgggcagcag cacggggccgtcgccgatggggg tgttctgctggtagtggtcggcgagctgcacgctgccgtcctcgatgttgtggcggatct tgaagttcaccttgatgccgttcttctg cttgtcggccatgatatagacgttgtggctgttgtagttgtactccagcttgtgccccag gatgttgccgtcctccttgaagtcgatgc ccttcagctcgatgcggttcaccagggtgtcgccctcgaacttcacctcggcgcgggtct tgtagttgccgtcgtccttgaagaag atggtgcgctcctggacgtagccttcgggcatggcggacttgaagaagtcgtgctgcttc atgtggtcggggtagcggctgaag cactgcacgccgtaggtcagggtggtcacgagggtgggccagggcacgggcagcttgccg gtggtgcagatgaacttcagg gtcagcttgccgtaggtggcatcgccctcgccctcgccggacacgctgaacttgtggccg tttacgtcgccgtccagctcgacca ggatgggcaccaccccggtgaacagctcctcgcccttgctcaccatggtggcgaccggta gcgctagcggatctgacggttca ctaaaccagctctgcttatatagacctcccaccgtacacgcctaccgcccatttgcgtca atggggcggagttgttacgacattttg gaaagtcccgttgattttggtgccaaaacaaactcccattgacgtcaatggggtggagac ttggaaatccccgtgagtcaaaccg ctatccacgcccattgatgtactgccaaaaccgcatcaccatggtaatagcgatgactaa tacgtagatgtactgccaagtaggaa agtcccataaggtcatgtactgggcataatgccaggcgggccatttaccgtcattgacgt caatagggggcgtacttggcatatga tacacttgatgtactgccaagtgggcagtttaccgtaaatactccacccattgacgtcaa tggaaagtccctattggcgttactatgg gaacatacgtcattattgacgtcaatgggcgggggtcgttgggcggtcagccaggcgggc catttaccgtaagttatgtaacgcg gaactccatatatgggctatgaactaatgaccccgtaattgattactattannnctagca gatctggtaccgtcgacgcggccgcg atatcctcgagaagctttctagagnnntaagggtgggaaagaatatataaggtgggggtc ttatgtagttttgtatctgttttgcagc agccgccgccgccatgagcaccaactcgtttgatggaagcattgtgagctcatatttgac aacgcgcatgcccccatgggccgg ggtgcgtcagaatgtgatgggctccagcattgatggtcgccccgtcctgcccgcaaactc tactaccttgacctacgagaccgtg tctggaacgccgttggagactgcagcctccgccgccgcttcagccgctgcagccaccgcc cgcgggattgtgactgactttgct ttcctgagcccgcttgcaagcagtgcagcttcccgttcatccgcccgcgatgacaagttg acggctcttttggcacaattggattctt tgacccgggaacttaatgtcgtttctcagcagctgttggatctgcgccagcaggtttctg ccctgaaggcttcctcccctcccaatg cggtttaaaacataaataaaaaaccagactctgtttggatttggatcaagcaagtgtctt gctgtctttatttaggggttttgcgcgcg cggtaggcccgggaccagcggtctcggtcgttgagggtcctgtgtattttttccaggacg tggtaaaggtgactctggatgttcag atacatgggcataagcccgtctctggggtggaggtagcaccactgcagagcttcatgctg cggggtggtgttgtagatgatccag tcgtagcaggagcgctgggcgtggtgcctaaaaatgtctttcagtagcaagctgattgcc aggggcaggcccttggtgtaagtgt ttacaaagcggttaagctgggatgggtgcatacgtggggatatgagatgcatcttggact gtatttttaggttggctatgttcccagc catatccctccggggattcatgttgtgcagaaccaccagcacagtgtatccggtgcactt gggaaatttgtcatgtagcttagaagg aaatgcgtggaagaacttggagacgcccttgtgacctccaagattttccatgcattcgtc cataatgatggcaatgggcccacgg gcggcggcctgggcgaagatatttctgggatcactaacgtcatagttgtgttccaggatg agatcgtcataggccatttttacaaag cgcgggcggagggtgccagactgcggtataatggttccatccggcccaggggcgtagtta ccctcacagatttgcatttcccac gctttgagttcagatggggggatcatgtctacctgcggggcgatgaagaaaacggtttcc ggggtaggggagatcagctggga agaaagcaggttcctgagcagctgcgacttaccgcagccggtgggcccgtaaatcacacc tattaccgggtgcaactggtagtt aagagagctgcagctgccgtcatccctgagcaggggggccacttcgttaagcatgtccct gactcgcatgttttccctgaccaaat ccgccagaaggcgctcgccgcccagcgatagcagttcttgcaaggaagcaaagtttttca acggtttgagaccgtccgccgtag gcatgcttttgagcgtttgaccaagcagttccaggcggtcccacagctcggtcacctgct ctacggcatctcgatccagcatatctc ctcgtttcgcgggttggggcggctttcgctgtacggcagtagtcggtgctcgtccagacg ggccagggtcatgtctttccacggg cgcagggtcctcgtcagcgtagtctgggtcacggtgaaggggtgcgctccgggctgcgcg ctggccagggtgcgcttgaggc tggtcctgctggtgctgaagcgctgccggtcttcgccctgcgcgtcggccaggtagcatt tgaccatggtgtcatagtccagccc ctccgcggcgtggcccttggcgcgcagcttgcccttggaggaggcgccgcacgaggggca gtgcagacttttgagggcgtag agcttgggcgcgagaaataccgattccggggagtaggcatccgcgccgcaggccccgcag acggtctcgcattccacgagcc aggtgagctctggccgttcggggtcaaaaaccaggtttcccccatgctttttgatgcgtt tcttacctctggtttccatgagccggtgt ccacgctcggtgacgaaaaggctgtccgtgtccccgtatacagactnnngtttaaacgaa ttcnnntataaaatgcaaggtgctg ctcaaaaaatcaggcaaagcctcgcgcaaaaaagaaagcacatcgtagtcatgctcatgc agataaaggcaggtaagctccgg aaccaccacagaaaaagacaccatttttctctcaaacatgtctgcgggtttctgcataaa cacaaaataaaataacaaaaaaacatt taaacattagaagcctgtcttacaacaggaaaaacaacccttataagcataagacggact acggccatgccggcgtgaccgtaa aaaaactggtcaccgtgattaaaaagcaccaccgacagctcctcggtcatgtccggagtc ataatgtaagactcggtaaacacat caggttgattcatcggtcagtgctaaaaagcgaccgaaatagcccgggggaatacatacc cgcaggcgtagagacaacattac agcccccataggaggtataacaaaattaataggagagaaaaacacataaacacctgaaaa accctcctgcctaggcaaaatagc accctcccgctccagaacaacatacagcgcttcacagcggcagcctaacagtcagcctta ccagtaaaaaagaaaacctattaa aaaaacaccactcgacacggcaccagctcaatcagtcacagtgtaaaaaagggccaagtg cagagcgagtatatataggacta aaaaatgacgtaacggttaaagtccacaaaaaacacccagaaaaccgcacgcgaacctac gcccagaaacgaaagccaaaa aacccacaacttcctcaaatcgtcacttccgttttcccacgttacgtaacttcccatttt aagaaaactacaattcccaacacatacaa gttactccgccctaaaacctacgtcacccgccccgttcccacgccccgcgccacgtcaca aactccaccccctcattatcatattg gcttcaatccaaaataaggtatattattgatgatnnnttaattaaggatccnnncggtgt gaaataccgcacagatgcgtaaggag aaaataccgcatcaggcgctcttccgcttcctcgctcactgactcgctgcgctcggtcgt tcggctgcggcgagcggtatcagct cactcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatg tgagcaaaaggccagcaaaagg ccaggaaccgtaaaaaggccgcgttgctggcgtttttccataggctccgcccccctgacg agcatcacaaaaatcgacgctcaa gtcagaggtggcgaaacccgacaggactataaagataccaggcgtttccccctggaagct ccctcgtgcgctctcctgttccga ccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtggcgctttctc atagctcacgctgtaggtatctcagttc ggtgtaggtcgttcgctccaagctgggctgtgtgcacgaaccccccgttcagcccgaccg ctgcgccttatccggtaactatcgt cttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaacagg attagcagagcgaggtatgtagg cggtgctacagagttcttgaagtggtggcctaactacggctacactagaaggacagtatt tggtatctgcgctctgctgaagccag ttaccttcggaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcg gtggtttttttgtttgcaagcagcaga ttacgcgcagaaaaaaaggatctcaagaagatcctttgatcttttctacggggtctgacg ctcagtggaacgaaaactcacgttaa gggattttggtcatgagattatcaaaaaggatcttcacctagatccttttaaattaaaaa tgaagttttaaatcaatctaaagtatatatg agtaaacttggtctgacagttaccaatgcttaatcagtgaggcacctatctcagcgatct gtctatttcgttcatccatagttgcctgac tccccgtcgtgtagataactacgatacgggagggcttaccatctggccccagtgctgcaa tgataccgcgagacccacgctcac cggctccagatttatcagcaataaaccagccagccggaagggccgagcgcagaagtggtc ctgcaactttatccgcctccatcc agtctattaattgttgccgggaagctagagtaagtagttcgccagttaatagtttgcgca acgttgttgnnnnnnaaaaaggatctt cacctagatccttttcacgtagaaagccagtccgcagaaacggtgctgaccccggatgaa tgtcagctactgggctatctggaca agggaaaacgcaagcgcaaagagaaagcaggtagcttgcagtgggcttacatggcgatag ctagactgggcggttttatggac agcaagcgaaccggaattgccagctggggcgccctctggtaaggttgggaagccctgcaa agtaaactggatggctttctcgc cgccaaggatctgatggcgcaggggatcaagctctgatcaagagacaggatgaggatcgt ttcgcatgattgaacaagatggat tgcacgcaggttctccggccgcttgggtggagaggctattcggctatgactgggcacaac agacaatcggctgctctgatgccg ccgtgttccggctgtcagcgcaggggcgcccggttctttttgtcaagaccgacctgtccg gtgccctgaatgaactgcaagacga ggcagcgcggctatcgtggctggccacgacgggcgttccttgcgcagctgtgctcgacgt tgtcactgaagcgggaagggact ggctgctattgggcgaagtgccggggcaggatctcctgtcatctcaccttgctcctgccg agaaagtatccatcatggctgatgca atgcggcggctgcatacgcttgatccggctacctgcccattcgaccaccaagcgaaacat cgcatcgagcgagcacgtactcg gatggaagccggtcttgtcgatcaggatgatctggacgaagagcatcaggggctcgcgcc agccgaactgttcgccaggctca aggcgagcatgcccgacggcgaggatctcgtcgtgacccatggcgatgcctgcttgccga atatcatggtggaaaatggccgc ttttctggattcatcgactgtggccggctgggtgtggcggaccgctatcaggacatagcg ttggctacccgtgatattgctgaaga gcttggcggcgaatgggctgaccgcttcctcgtgctttacggtatcgccgctcccgattc gcagcgcatcgccttctatcgccttct tgacgagttcttctgaattttgttaaaatttttgttaaatcagctcattttttaaccaat aggccgaaatcggcaacatcccttataaatca aaagaatagaccgcgatagggttgagtgttgttccagtttggaacaagagtccactatta aagaacgtggactccaacgtcaaag ggcgaaaaaccgtctatcagggcgatggcccactacgtgaaccatcacccaaatcaagtt ttttgcggtcgaggtgccgtaaag ctctaaatcggaaccctaaagggagcccccgatttagagcttgacggggaaagccggcga acgtggcgagaaaggaaggga agaaagcgaaaggagcgggcgctagggcgctggcaagtgtagcggtcacgctgcgcgtaa ccaccacacccgcgcgcttaa tgcgccg pAdTrack-CMV Sequencel90476 Adenovirus Packaging or Helper (SEQ ID NO:

9) tgttaacttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaattt cacaaataaagcatttttttcactgcattcta gttgtggtttgtccaaactcatcaatgtatcttaacgcggaggtttatcgacgatctgct agtgattaatagtaatcaattacggggtc attagttcatagcccatatatggagttccgcgttacataacttacggtaaatggcccgcc tggctgaccgcccaacgacccccgcc cattgacgtcaataatgacgtatgttcccatagtaacgccaatagggactttccattgac gtcaatgggtggagtatttacggtaaac tgcccacttggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaa tgacggtaaatggcccgcctggcatt atgcccagtacatgaccttatgggactttcctacttggcagtacatctacgtattagtca tcgctattaccatggtgatgcggttttggc agtacatcaatgggcgtggatagcggtttgactcacggggatttccaagtctccacccca ttgacgtcaatgggagtttgttttggc accaaaatcaacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgg gcggtaggcgtgtacggtgggag gtctatataagcagagctggtttagtgaaccgtcagatccgctagcgctaccggtcgcca ccatggtgagcaagggcgaggag ctgttcaccggggtggtgcccatcctggtcgagctggacggcgacgtaaacggccacaag ttcagcgtgtccggcgagggcg agggcgatgccacctacggcaagctgaccctgaagttcatctgcaccaccggcaagctgc ccgtgccctggcccaccctcgtg accaccctgacctacggcgtgcagtgcttcagccgctaccccgaccacatgaagcagcac gacttcttcaagtccgccatgccc gaaggctacgtccaggagcgcaccatcttcttcaaggacgacggcaactacaagacccgc gccgaggtgaagttcgagggcg acaccctggtgaaccgcatcgagctgaagggcatcgacttcaaggaggacggcaacatcc tggggcacaagctggagtacaa ctacaacagccacaacgtctatatcatggccgacaagcagaagaacggcatcaaggtgaa cttcaagatccgccacaacatcg aggacggcagcgtgcagctcgccgaccactaccagcagaacacccccatcggcgacggcc ccgtgctgctgcccgacaacc actacctgagcacccagtccgccctgagcaaagaccccaacgagaagcgcgatcacatgg tcctgctggagttcgtgaccgcc gccgggatcactctcggcatggacgagctgtacaagtccggactcagatccaccggatct agataactgatcataatcagccata ccacatttgtagaggttttacttgctttaaaaaacctcccacacctccccctgaacctga aacataaaatgaatgcaattgttgttgtta acttgtttattgcagcttataatggttacaaataaagcaatagcatcacaaatttcacaa ataaagcatttttttcactgcattctagttgt ggtttgtccaaactcatcaatgtatcttaacgcgatcaagctagcttgctagactcgact gactataataataaaacgccaactttga cccggaacgcggaaaacacctgagaaaaacacctgggcgagtctccacgtaaacggtcaa agtccccgcggccctagacaa atattacgcgctatgagtaacacaaaattattcagatttcacttcctcttattcagtttt cccgcgaaaatggccaaatcttactcggtta cgcccaaatttactacaacatccgcctaaaaccgcgcgaaaattgtcacttcctgtgtac accggcgcacaccaaaaacgtcactt ttgccacatccgtcgcttacatgtgttccgccacacttgcaacatcacacttccgccaca ctactacgtcacccgccccgttcccac gccccgcgccacgtcacaaactccaccccctcattatcatattggcttcaatccaaaata aggtatattattgatgatgttaattaaga attaattcgatcctgaatggcgaatggacgcgccctgtagcggcgcattaagcgcggcgg gtgtggtggttacgcgcagcgtga ccgctacacttgccagcgccctagcgcccgctcctttcgctttcttcccttcctttctcg ccacgttcgccggctttccccgtcaagct ctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccccaaa aaacttgattagggtgatggttcacgt agtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttcttt aatagtggactcttgttccaaactggaac aacactcaaccctatctcggtctattcttttgatttataagggattttgccgatttcggc ctattggttaaaaaatgagctgatttaacaa aaattttaacaaaattcagaagaactcgtcaagaaggcgatagaaggcgatgcgctgcga atcgggagcggcgataccgtaaa gcacgaggaagcggtcagcccattcgccgccaagctcttcagcaatatcacgggtagcca acgctatgtcctgatagcggtccg ccacacccagccggccacagtcgatgaatccagaaaagcggccattttccaccatgatat tcggcaagcaggcatcgccatgg gtcacgacgagatcctcgccgtcgggcatgctcgccttgagcctggcgaacagttcggct ggcgcgagcccctgatgctcttcg tccagatcatcctgatcgacaagaccggcttccatccgagtacgtgctcgctcgatgcga tgtttcgcttggtggtcgaatgggca ggtagccggatcaagcgtatgcagccgccgcattgcatcagccatgatggatactttctc ggcaggagcaaggtgagatgaca ggagatcctgccccggcacttcgcccaatagcagccagtcccttcccgcttcagtgacaa cgtcgagcacagctgcgcaagga acgcccgtcgtggccagccacgatagccgcgctgcctcgtcttgcagttcattcagggca ccggacaggtcggtcttgacaaaa agaaccgggcgcccctgcgctgacagccggaacacggcggcatcagagcagccgattgtc tgttgtgcccagtcatagccga atagcctctccacccaagcggccggagaacctgcgtgcaatccatcttgttcaatcatgc gaaacgatcctcatcctgtctcttgat cagagcttgatcccctgcgccatcagatccttggcggcaagaaagccatccagtttactt tgcagggcttcccaaccttaccagag ggcgccccagctggcaattccggttcgcttgctgtccataaaaccgcccagtctagctat cgccatgtaagcccactgcaagcta cctgctttctctttgcgcttgcgttttcccttgtccagatagcccagtagctgacattca tccggggtcagcaccgtttctgcggactg gctttctacgtgaaaaggatctaggtgaagatcctttttgataatctcatggctgcagca atggcaacaacgttgcgcaaactattaa ctggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggata aagttgcaggaccacttctgcgctcg gcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgc ggtatcattgcagcactggggccag atggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatg aacgaaatagacagatcgctgagat aggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatacttta gattgatttaaaacttcatttttaatttaaaa ggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagtttt cgttccactgagcgtcagaccccgtaga aaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaac aaaaaaaccaccgctaccagcggtggtt tgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcg cagataccaaatactgtccttctagtgt agccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgc taatcctgttaccagtggctgctgcca gtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgc agcggtcgggctgaacggggggt tcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgt gagctatgagaaagcgccacgct tcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcg cacgagggagcttccaggg ggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcga tttttgtgatgctcgtcaggggggcgg agcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcct tttgctcacatgttctttcctgcgttatcc cctgattctgtggataaccgtattaccgcctttgagtgagctgataccgctcgccgcagc cgaacgaccgagcgcagcgagtca gtgagcgaggaagcggaagagcgcctgatgcggtattttctccttacgcatctgtgcggt atttcacaccgcatatggatccatgc atgttaattaacatcatcaataatataccttattttggattgaagccaatatgataatga gggggtggagtttgtgacgtggcgcggg gcgtgggaacggggcgggtgacgtaggttttagggcggagtaacttgtatgtgttgggaa ttgtagttttcttaaaatgggaagtg acgtaacgtgggaaaacggaagtgacgatttgaggaagttgtgggttttttggctttcgt ttctgggcgtaggttcgcgtgcggtttt ctgggtgttttttgtggactttaaccgttacgtcattttttagtcctatatatactcgct ctgcacttggcccttttttacactgtgactgatt gagctggtgccgtgtcgagtggtgtttttttaataggttttcttttttactggtaaggct gactgttatggctgccgctgtggaagcgct gtatgttgttctggagcgggagggtgctattttgcctaggcaggagggtttttcaggtgt ttatgtgtttttctctcctattaattttgttat acctcctatgggggctgtaatgttgtctctacgcctgcgggtatgtattcccccgggcta tttcggtcgctttttagcactgaccgatg tgaatcaacctgatgtgtttaccgagtcttacattatgactccggacatgaccgaggagc tgtcggtggtgctttttaatcacggtga ccagtttttttacggtcacgccggcatggccgtagtccgtcttatgcttataagggttgt ttttcctgttgtaagacaggcttctaatgttt aaatgtttttttgttattttattttgtgtttatgcagaaacccgcagacatgtttgagag aaaaatggtgtctttttctgtggtggttccgga gcttacctgcctttatctgcatgagcatgactacgatgtgctttcttttttgcgcgaggc tttgcctgattttttgagcagcaccttgcatt ttatatcgccgcccatgcaacaagctattgaattcgtttaaactccctctcaagtctgta tacggggacacggacagccttttcgtca ccgagcgtggacaccggctcatggaaaccagaggtaagaaacgcatcaaaaagcatgggg gaaacctggtttttgaccccga acggccagagctcacctggctcgtggaatgcgagaccgtctgcggggcctgcggcgcgga tgcctactccccggaatcggta tttctcgcgcccaagctctacgccctcaaaagtctgcactgcccctcgtgcggcgcctcc tccaagggcaagctgcgcgccaag ggccacgccgcggaggggctggactatgacaccatggtcaaatgctacctggccgacgcg cagggcgaagaccggcagcg cttcagcaccagcaggaccagcctcaagcgcaccctggccagcgcgcagcccggagcgca ccccttcaccgtgacccagac tacgctgacgaggaccctgcgcccgtggaaagacatgaccctggcccgtctggacgagca ccgactactgccgtacagcgaa agccgccccaacccgcgaaacgaggagatatgctggatcgagatgccgtagagcaggtga ccgagctgtgggaccgcctgg aactgcttggtcaaacgctcaaaagcatgcctacggcggacggtctcaaaccgttgaaaa actttgcttccttgcaagaactgcta tcgctgggcggcgagcgccttctggcggatttggtcagggaaaacatgcgagtcagggac atgcttaacgaagtggcccccct gctcagggatgacggcagctgcagctctcttaactaccagttgcagccggtaataggtgt gatttacgggcccaccggctgcggt aagtcgcagctgctcaggaacctgctttcttcccagctgatctcccctaccccggaaacc gttttcttcatcgccccgcaggtagac atgatccccccatctgaactcaaagcgtgggaaatgcaaatctgtgagggtaactacgcc cctgggccggatggaaccattatac cgcagtctggcaccctccgcccgcgctttgtaaaaatggcctatgacgatctcatcctgg aacacaactatgacgttagtgatccc agaaatatcttcgcccaggccgccgcccgtgggcccattgccatcattatggacgaatgc atggaaaatcttggaggtcacaag ggcgtctccaagttcttccacgcatttccttctaagctacatgacaaatttcccaagtgc accggatacactgtgctggtggttctgc acaacatgaatccccggagggatatggctgggaacatagccaacctaaaaatacagtcca agatgcatctcatatccccacgtat gcacccatcccagcttaaccgctttgtaaacacttacaccaagggcctgcccctggcaat cagcttgctactgaaagacatttttag gcaccacgcccagcgctcctgctacgactggatcatctacaacaccaccccgcagcatga agctctgcagtggtgctacctcca ccccagagacgggcttatgcccatgtatctgaacatccagagtcacctttaccacgtcct ggaaaaaatacacaggaccctcaac gaccgagaccgctggtcccgggcctaccgcgcgcgcaaaacccctaaataaagacagcaa gacacttgcttgatccaaatcca aacagagtctggttttttatttatgttttaaaccgcattgggaggggaggaagccttcag ggcagaaacctgctggcgcagatcca acagctgctgagaaacgacattaagttcccgggtcaaagaatccaattgtgccaaaagag ccgtcaacttgtcatcgcgggcgg atgaacgggaagctgcactgcttgcaagcgggctcaggaaagcaaagtcagtcacaatcc cgcgggcggtggctgcagcgg ctgaagcggcggcggaggctgcagtctccaacggcgttccagacacggtctcgtaggtca aggtagtagagtttgcgggcag gacggggcgaccatcaatgctggagcccatcacattctgacgcaccccggcccatggggg catgcgcgttgtcaaatatgagc tcacaatgcttccatcaaacgagttggtgctcatggcggcggcggctgctgcaaaacaga tacaaaactacataagacccccac cttatatattctttcccacccttaaccacgcccagatcctctagcagtgataaacgtcta atagtaatcaattacggggtcattagttca tagcccatatatggagttccgcgttacataacttacggtaaatggcccgcctggctgacc gcccaacgacccccgcccattgacg tcaataatgacgtatgttcccatagtaacgccaatagggactttccattgacgtcaatgg gtggagtatttacggtaaactgcccact tggcagtacatcaagtgtatcatatgccaagtacgccccctattgacgtcaatgacggta aatggcccgcctggcattatgcccag tacatgaccttatgggactttcctacttggcagtacatctacgtattagtcatcgctatt accatggtgatgcggttttggcagtacatc aatgggcgtggatagcggtttgactcacggggatttccaagtctccaccccattgacgtc aatgggagtttgttttggcaccaaaat caacgggactttccaaaatgtcgtaacaactccgccccattgacgcaaatgggcggtagg cgtgtacggtgggaggtctatataa gcagagctggtttagtgaaccgtcagatccgctagagatctggtaccgtcgacgcggccg ctcgagcctaagcttctagataag atatccgatccaccggatctagataactgatcataatcagccataccacatttgtagagg ttttacttgctttaaaaaacctcccacac ctccccctgaacctgaaacataaaatgaatgcaattgttgt pZB3 AAV dSaCas9 One Vector (SEQ ID NO: 10) aaaacctctgacacatgcagctcccggagacggtcacagcttgtctgtaagcggatgccg ggagcagacaagcccgtcaggg cgcgtcagcgggtgttggcgggtgtcggggctggcttaactatgcggcatcagagcagat tgtactgagagtgcaccataaaatt gtaaacgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcatttttt aaccaatagaccgaaatcggcaaaatcccttat aaatcaaaagaatagcccgagatagagttgagtgttgttccagtttggaacaagagtcca ctattaaagaacgtggactccaacgt caaagggcgaaaaaccgtctatcagggcgatggcccactacgtgaaccatcacccaaatc aagttttttggggtcgaggtgccg taaagcactaaatcggaaccctaaagggagcccccgatttagagcttgacggggaaagcc ggcgaacgtggcgagaaagga agggaagaaagcgaaaggagcgggcgctaaggcgctggcaagtgtagcggtcacgctgcg cgtaaccaccacacccgccg cgcttaatgcgccgctacagggcgcgtactatggttgctttgacgtatgcggtgtgaaat accgcacagatgcgtaaggagaaaa taccgcatcaggcgcccctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgg gcaaagcccgggcgtcgggcg acctttggtcgcccggcctcagtgagcgagcgagcgcgcagagagggagtggccaactcc atcactaggggttcctgcggcc gcgagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagag agataattggaattaatttgactgtaaac acaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgca gttttaaaattatgttttaaaatggactatc atatgcttaccgtaacttgaaagtatttcgatttcttggctttatatatcttgtggaaag gacgaaacaccgggagaccacggcaggt ctcagttttagtactctggaggcc AAC AT GAGGAT C ACCC ATGTCTGC AGggccaacagaatctactaaa acaaggcaaaatgccgtgtttatctcgtcaacttgttggcgagatttttttggccgcctc gaggtacttatataagggggtgggggc gcgttcgtcctcagtcgcgatcgaacactcgagccgagcagacgtgcctacggaccgacc ggtgccaccatggccccaaaga agaagcggaaggtcggtatccacggagtcccagcagccaagcggaactacatcctgggcc tggccatcggcatcaccagcgt gggctacggcatcatcgactacgagacacgggacgtgatcgatgccggcgtgcggctgtt caaagaggccaacgtggaaaac aacgagggcaggcggagcaagagaggcgccagaaggctgaagcggcggaggcggcataga atccagagagtgaagaag ctgctgttcgactacaacctgctgaccgaccacagcgagctgagcggcatcaacccctac gaggccagagtgaagggcctga gccagaagctgagcgaggaagagttctctgccgccctgctgcacctggccaagagaagag gcgtgcacaacgtgaacgagg tggaagaggacaccggcaacgagctgtccaccaaagagcagatcagccggaacagcaagg ccctggaagagaaatacgtg gccgaactgcagctggaacggctgaagaaagacggcgaagtgcggggcagcatcaacaga ttcaagaccagcgactacgtg aaagaagccaaacagctgctgaaggtgcagaaggcctaccaccagctggaccagagcttc atcgacacctacatcgacctgct ggaaacccggcggacctactatgagggacctggcgagggcagccccttcggctggaagga catcaaagaatggtacgagat gctgatgggccactgcacctacttccccgaggaactgcggagcgtgaagtacgcctacaa cgccgacctgtacaacgccctga acgacctgaacaatctcgtgatcaccagggacgagaacgagaagctggaatattacgaga agttccagatcatcgagaacgtgt tcaagcagaagaagaagcccaccctgaagcagatcgccaaagaaatcctcgtgaacgaag aggatattaagggctacagagt gaccagcaccggcaagcccgagttcaccaacctgaaggtgtaccacgacatcaaggacat taccgcccggaaagagattattg agaacgccgagctgctggatcagattgccaagatcctgaccatctaccagagcagcgagg acatccaggaagaactgaccaat ctgaactccgagctgacccaggaagagatcgagcagatctctaatctgaagggctatacc ggcacccacaacctgagcctgaa ggccatcaacctgatcctggacgagctgtggcacaccaacgacaaccagatcgctatctt caaccggctgaagctggtgcccaa gaaggtggacctgtcccagcagaaagagatccccaccaccctggtggacgacttcatcct gagccccgtcgtgaagagaagct tcatccagagcatcaaagtgatcaacgccatcatcaagaagtacggcctgcccaacgaca tcattatcgagctggcccgcgaga agaactccaaggacgcccagaaaatgatcaacgagatgcagaagcggaaccggcagacca acgagcggatcgaggaaatc atccggaccaccggcaaagagaacgccaagtacctgatcgagaagatcaagctgcacgac atgcaggaaggcaagtgcctgt acagcctggaagccatccctctggaagatctgctgaacaaccccttcaactatgaggtgg accacatcatccccagaagcgtgtc cttcgacaacagcttcaacaacaaggtgctcgtgaagcaggaagaagccagcaagaaggg caaccggaccccattccagtac ctgagcagcagcgacagcaagatcagctacgaaaccttcaagaagcacatcctgaatctg gccaagggcaagggcagaatca gcaagaccaagaaagagtatctgctggaagaacgggacatcaacaggttctccgtgcaga aagacttcatcaaccggaacctg gtggataccagatacgccaccagaggcctgatgaacctgctgcggagctacttcagagtg aacaacctggacgtgaaagtgaa gtccatcaatggcggcttcaccagctttctgcggcggaagtggaagtttaagaaagagcg gaacaaggggtacaagcaccacg ccgaggacgccctgatcattgccaacgccgatttcatcttcaaagagtggaagaaactgg acaaggccaaaaaagtgatggaa aaccagatgttcgaggaaaagcaggccgagagcatgcccgagatcgaaaccgagcaggag tacaaagagatcttcatcaccc cccaccagatcaagcacattaaggacttcaaggactacaagtacagccaccgggtggaca agaagcctaatagagagctgatt aacgacaccctgtactccacccggaaggacgacaagggcaacaccctgatcgtgaacaat ctgaacggcctgtacgacaagg acaatgacaagctgaaaaagctgatcaacaagagccccgaaaagctgctgatgtaccacc acgacccccagacctaccagaa actgaagctgattatggaacagtacggcgacgagaagaatcccctgtacaagtactacga ggaaaccgggaactacctgacca agtactccaaaaaggacaacggccccgtgatcaagaagattaagtattacggcaacaaac tgaacgcccatctggacatcaccg acgactaccccaacagcagaaacaaggtcgtgaagctgtccctgaagccctacagattcg acgtgtacctggacaatggcgtgt acaagttcgtgaccgtgaagaatctggatgtgatcaaaaaagaaaactactacgaagtga atagcaagtgctatgaggaagctaa gaagctgaagaagatcagcaaccaggccgagtttatcgcctccttctacaacaacgatct gatcaagatcaacggcgagctgtat agagtgatcggcgtgaacaacgacctgctgaaccggatcgaagtgaacatgatcgacatc acctaccgcgagtacctggaaaa catgaacgacaagaggccccccaggatcattaagacaatcgcctccaagacccagagcat taagaagtacagcacagacattc tgggcaacctgtatgaagtgaaatctaagaagcaccctcagatcatcaaaaagggcaaaa ggccggcggccacgaaaaaggc cggccaggcaaaaaagaaaaagggatccgaggccagcggttccggacgggctgacgcatt ggacgattttgatctggatatgc tgggaagtgacgccctcgatgattttgaccttgacatgcttggttcggatgcccttgatg actttgacctcgacatgctcggcagtg acgcccttgatgatttcgacctggacatgctgattaactctagaagttccggatctccga aaaagaaacgcaaagttggttcggga ggtggttcgggtggctctggatcagtgctgcctcaggctcctgctcctgcaccagctcca gccatggtgtctgcactggctcagg caccagcacccgtgcctgtgctggctcctggacctccacaggctgtggctccaccagccc ctaaacctacacaggccggcgag ggcacactgtctgaagctctgctgcagctgcagttcgacgacgaggatctgggagccctg ctgggaaacagcaccgatcctgc cgtgttcaccgacctggccagcgtggacaacagcgagttccagcagctgctgaaccaggg catccctgtggcccctcacacca ccgagcccatgctgatggaataccccgaggccatcacccggctcgtgacaggcgctcaga ggcctcctgatccagctcctgcc cctctgggagcaccaggcctgcctaatggactgctgtctggcgacgaggacttcagctct atcgccgatatggatttctcagcctt gctgtcaggcggtggtagtggtgggagcggtagtgacctttcccatccgcccccaagggg ccatctggatgagctgacaacca cacttgagtccatgaccgaggatctgaacctggactcacccctgaccccggaattgaacg agattctggataccttcctgaacga cgagtgcctcttgcatgccatgcatatcagcacaggactgtccatcttcgacacatctct gttttaggaattcaaataaaatacgaaa tgaaataaaatacgaaatgcaattgggccgcaggaacccctagtgatggagttggccact ccctctctgcgcgctcgctcgctca ctgaggccgggcgaccaaaggtcgcccgacgcccgggctttgcccgagcggcctcagtga gcgagcgagcgcgcagctgc ctgcaggacatgtgagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttg ctggcgtttttccataggctccg cccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaaacccgacagg actataaagataccaggcgtttc cccctggaagctccctcgtgcgctctcctgttccgaccctgccgcttaccggatacctgt ccgcctttctcccttcgggaagcgtg gcgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaag ctgggctgtgtgcacgaaccccccgtt cagcccgaccgctgcgccttatccggtaactatcgtcttgagtccaacccggtaagacac gacttatcgccactggcagcagcc actggtaacaggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtgg tggcctaactacggctacactagaa gaacagtatttggtatctgcgctctgctgaagccagttaccttcggaaaaagagttggta gctcttgatccggcaaacaaaccacc gctggtagcggtggtttttttgtttgcaagcagcagattacgcgcagaaaaaaaggatct caagaagatcctttgatcttttctacgg ggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatgagattatcaa aaaggatcttcacctagatccttttaaa ttaaaaatgaagttttaaatcaatctaaagtatatatgagtaaacttggtctgacagtta ccaatgcttaatcagtgaggcacctatctc agcgatctgtctatttcgttcatccatagttgcctgactccccgtcgtgtagataactac gatacgggagggcttaccatctggcccc agtgctgcaatgataccgcggcttccacgctcaccggctccagatttatcagcaataaac cagccagccggaagggccgagcg cagaagtggtcctgcaactttatccgcctccatccagtctattaattgttgccgggaagc tagagtaagtagttcgccagttaatagt ttgcgcaacgttgttgccattgctacaggcatcgtggtgtcacgctcgtcgtttggtatg gcttcattcagctccggttcccaacgat caaggcgagttacatgatcccccatgttgtgcaaaaaagcggttagctccttcggtcctc cgatcgttgtcagaagtaagttggcc gcagtgttatcactcatggttatggcagcactgcataattctcttactgtcatgccatcc gtaagatgcttttctgtgactggtgagtac tcaaccaagtcattctgagaatagtgtatgcggcgaccgagttgctcttgcccggcgtca atacgggataataccgcgccacata gcagaactttaaaagtgctcatcattggaaaacgttcttcggggcgaaaactctcaagga tcttaccgctgttgagatccagttcga tgtaacccactcgtgcacccaactgatcttcagcatcttttactttcaccagcgtttctg ggtgagcaaaaacaggaaggcaaaatg ccgcaaaaaagggaataagggcgacacggaaatgttgaatactcatactcttcctttttc aatattattgaagcatttatcagggttat tgtctcatgagcggatacatatttgaatgtatttagaaaaataaacaaataggggttccg cgcacatttccccgaaaagtgccacct gacgtctaagaaaccattattatcatgacattaacctataaaaataggcgtatcacgagg ccctttcgtctcgcgcgtttcggtgatg acggtg pGW047 AAV dSaCas9 Two Vector (SEQ ID NO: 11) cctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtc gggcgacctttggtcgcccgg cctcagtgagcgagcgagcgcgcagagagggagtggccaactccatcactaggggttcct gcggccgcacgcgttctagaaa gagggcctatttcccatgattccttcatatttgcatatacgatacaaggctgttagagag ataattggaattaatttgactgtaaacaca aagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggtagtttgcagtt ttaaaattatgttttaaaatggactatcatat gcttaccgtaacttgaaagtatttcgatttcttggctttatatatcttgtggaaaggacg aaacaccggaagagcgagctcttctgtttt agtactctggaggccaacatgaggatcacccatgtctgcagggccaacagaatctactaa aacaaggcaaaatgccgtgtttatc tcgtcaactggccaacatgaggatcacccatgtctgcagggcctgttggcgagatttttt tgctagctgcaaagatggataaagtttt aaacagagaggaatctttgcagctaatggaccttctaggtcttgaaaggagtgggaattg gctccggtgcccgtcagtgggcag agcgcacatcgcccacagtccccgagaagttggggggaggggtcggcaattgaaccggtg cctagagaaggtggcgcggg gtaaactgggaaagtgatgtcgtgtactggctccgcctttttcccgagggtgggggagaa ccgtatataagtgcagtagtcgccg tgaacgttctttttcgcaacgggtttgccgccagaacacaggtaagtgccgtgtgtggtt cccgcgggcctggcctctttacgggtt atggcccttgcgtgccttgaattacttccacctggctgcagtacgtgattcttgatcccg agcttcgggttggaagtgggtgggaga gttcgaggccttgcgcttaaggagccccttcgcctcgtgcttgagttgaggcctggcctg ggcgctggggccgccgcgtgcgaa tctggtggcaccttcgcgcctgtctcgctgctttcgataagtctctagccatttaaaatt tttgatgacctgctgcgacgctttttttctg gcaagatagtcttgtaaatgcgggccaagatctgcacactggtatttcggtttttggggc cgcgggcggcgacggggcccgtgc gtcccagcgcacatgttcggcgaggcggggcctgcgagcgcggccaccgagaatcggacg ggggtagtctcaagctggcc ggcctgctctggtgcctggcctcgcgccgccgtgtatcgccccgccctgggcggcaaggc tggcccggtcggcaccagttgc gtgagcggaaagatggccgcttcccggccctgctgcagggagctcaaaatgaaggacgcg gcgctcgggagagcgggcgg gtgagtcacccacacaaaggaaaagggcctttccgtcctcagccgtcgcttcatgtgact ccacggagtaccgggcgccgtcca ggcacctcgattagttctcgagcttttggagtacgtcgtctttaggttggggggaggggt tttatgcgatggagtttccccacactga gtgggtggagactgaagttaggccagcttggcacttgatgtaattctccttggaatttgc cctttttgagtttggatcttggttcattctc aagcctcagacagtggttcaaagtttttttcttccatttcaggtgtcgtgacgtacggcc accatggcttcaaactttactcagttcgtg ctcgtggacaatggtgggacaggggatgtgacagtggctccttctaatttcgctaatggg gtggcagagtggatcagctccaact cacggagccaggcctacaaggtgacatgcagcgtcaggcagtctagtgcccagaagagaa agtataccatcaaggtggaggt ccccaaagtggctacccagacagtgggcggagtcgaactgcctgtcgccgcttggaggtc ctacctgaacatggagctcactat cccaattttcgctaccaattctgactgtgaactcatcgtgaaggcaatgcaggggctcct caaagacggtaatcctatcccttccgc catcgccgctaactcaggtatctacagcgctggaggaggtggaagcggaggaggaggaag cggaggaggaggtagcggac ctaagaaaaagaggaaggtggcggccgctggatccccttcagggcagatcagcaaccagg ccctggctctggcccctagctc cgctccagtgctggcccagactatggtgccctctagtgctatggtgcctctggcccagcc acctgctccagcccctgtgctgacc ccaggaccaccccagtcactgagcgctccagtgcccaagtctacacaggccggcgagggg actctgagtgaagctctgctgc acctgcagttcgacgctgatgaggacctgggagctctgctggggaacagcaccgatcccg gagtgttcacagatctggcctcc gtggacaactctgagtttcagcagctgctgaatcagggcgtgtccatgtctcatagtaca gccgaaccaatgctgatggagtacc ccgaagccattacccggctggtgaccggcagccagcggccccccgaccccgctccaactc ccctgggaaccagcggcctgc ctaatgggctgtccggagatgaagacttctcaagcatcgctgatatggactttagtgccc tgctgtcacagatttcctctagtgggc agggaggaggtggaagcggcttcagcgtggacaccagtgccctgctggacctgttcagcc cctcggtgaccgtgcccgacat gagcctgcctgaccttgacagcagcctggccagtatccaagagctcctgtctccccagga gccccccaggcctcccgaggcag agaacagcagcccggattcagggaagcagctggtgcactacacagcgcagccgctgttcc tgctggaccccggctccgtgga caccgggagcaacgacctgccggtgctgtttgagctgggagagggctcctacttctccga aggggacggcttcgccgaggac cccaccatctccctgctgacaggctcggagcctcccaaagccaaggaccccactgtctcc taagaattcacgcgttaagtcgac aatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgct ccttttacgctatgtggatacgctgctttaat gcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttgtataaatc ctggttgctgtctctttatgaggagttgtggcc cgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttg gggcattgccaccacctgtcagctc ctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccgcctgc cttgcccgctgctggacaggggctcg gctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttccttggct gctcgcctgtgttgccacctggattctg cgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcccgc ggcctgctgccggctctgcggcctct tccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctccccgcg tcgactttaagaccaatgacgtgcgg accgagcggccgcaggaacccctagtgatggagttggccactccctctctgcgcgctcgc tcgctcactgaggccgggcgac caaaggtcgcccgacgcccgggctttgcccgggcggcctcagtgagcgagcgagcgcgca gctgcctgcaggggcgcctg atgcggtattttctccttacgcatctgtgcggtatttcacaccgcatacgtcaaagcaac catagtacgcgccctgtagcggcgcat taagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctag cgcccgctcctttcgctttcttccct tcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctcccttta gggttccgatttagtgctttacggcacct cgaccccaaaaaacttgatttgggtgatggttcacgtagtgggccatcgccctgatagac ggtttttcgccctttgacgttggagtc cacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcggg ctattcttttgatttataagggattttgccg atttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaac aaaatattaacgtttacaattttatggtgca ctctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacac ccgctgacgcgccctgacgggctt gtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtc agaggttttcaccgtcatcaccgaaa cgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataata atggtttcttagacgtcaggtggcactt ttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgt atccgctcatgagacaataaccctgataaa tgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgccctta ttcccttttttgcggcattttgccttcctgttt ttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgag tgggttacatcgaactggatctcaa cagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcacttt taaagttctgctatgtggcgcggtattat cccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgact tggttgagtactcaccagtcacaga aaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgag tgataacactgcggccaacttacttc tgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatg taactcgccttgatcgttgggaacc ggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggc aacaacgttgcgcaaactattaa ctggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggata aagttgcaggaccacttctgcgctcg gcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgc ggtatcattgcagcactggggccag atggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatg aacgaaatagacagatcgctgagat aggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatacttta gattgatttaaaacttcatttttaatttaaaa ggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagtttt cgttccactgagcgtcagaccccgtaga aaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaac aaaaaaaccaccgctaccagcggtggtt tgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcg cagataccaaatactgtccttctagtgt agccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgc taatcctgttaccagtggctgctgcca gtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgc agcggtcgggctgaacggggggt tcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgt gagctatgagaaagcgccacgct tcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcg cacgagggagcttccaggg ggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcga tttttgtgatgctcgtcaggggggcgg agcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcct tttgctcacatgt pGW060 AAV dSaCas9 Two Vector (SEQ ID NO: 12) cctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgggcgtcgggcgaccttt ggtcgcccggcctcagtgagcg agcgagcgcgcagagagggagtggccaactccatcactaggggttcctgcggcctctaga aaggatctgcgatcgctccggt gcccgtcagtgggcagagcgcacatcgcccacagtccccgagaagttggggggaggggtc ggcaattgaacgggtgcctag agaaggtggcgcggggtaaactgggaaagtgatgtcgtgtactggctccgcctttttccc gagggtgggggagaaccgtatata agtgcagtagtcgccgtgaacgttctttttcgcaacgggtttgccgccagaacacagctg aagcttcgaggggctcgcatctctcc ttcacgcgcccgccgccctacctgaggccgccatccacgccggttgagtcgcgttctgcc gcctcccgcctgtggtgcctcctg aactgcgtccgccgtctaggtaagtttaaagctcaggtcgagaccgggcctttgtccggc gctcccttggagcctacctagactc agccggctctccacgctttgcctgaccctgcttgctcaactctacgtctttgtttcgttt tctgttctgcgccgttacagatctggctaac taccggtgccaccatggccccaaagaagaagcggaaggtcggtatccacggagtcccagc agccaagcggaactacatcctg ggcctggccatcggcatcaccagcgtgggctacggcatcatcgactacgagacacgggac gtgatcgatgccggcgtgcggc tgttcaaagaggccaacgtggaaaacaacgagggcaggcggagcaagagaggcgccagaa ggctgaagcggcggaggcg gcatagaatccagagagtgaagaagctgctgttcgactacaacctgctgaccgaccacag cgagctgagcggcatcaacccct acgaggccagagtgaagggcctgagccagaagctgagcgaggaagagttctctgccgccc tgctgcacctggccaagagaa gaggcgtgcacaacgtgaacgaggtggaagaggacaccggcaacgagctgtccaccaaag agcagatcagccggaacagc aaggccctggaagagaaatacgtggccgaactgcagctggaacggctgaagaaagacggc gaagtgcggggcagcatcaa cagattcaagaccagcgactacgtgaaagaagccaaacagctgctgaaggtgcagaaggc ctaccaccagctggaccagag cttcatcgacacctacatcgacctgctggaaacccggcggacctactatgagggacctgg cgagggcagccccttcggctgga aggacatcaaagaatggtacgagatgctgatgggccactgcacctacttccccgaggaac tgcggagcgtgaagtacgcctac aacgccgacctgtacaacgccctgaacgacctgaacaatctcgtgatcaccagggacgag aacgagaagctggaatattacga gaagttccagatcatcgagaacgtgttcaagcagaagaagaagcccaccctgaagcagat cgccaaagaaatcctcgtgaacg aagaggatattaagggctacagagtgaccagcaccggcaagcccgagttcaccaacctga aggtgtaccacgacatcaagga cattaccgcccggaaagagattattgagaacgccgagctgctggatcagattgccaagat cctgaccatctaccagagcagcga ggacatccaggaagaactgaccaatctgaactccgagctgacccaggaagagatcgagca gatctctaatctgaagggctata ccggcacccacaacctgagcctgaaggccatcaacctgatcctggacgagctgtggcaca ccaacgacaaccagatcgctatc ttcaaccggctgaagctggtgcccaagaaggtggacctgtcccagcagaaagagatcccc accaccctggtggacgacttcat cctgagccccgtcgtgaagagaagcttcatccagagcatcaaagtgatcaacgccatcat caagaagtacggcctgcccaacg acatcattatcgagctggcccgcgagaagaactccaaggacgcccagaaaatgatcaacg agatgcagaagcggaaccggc agaccaacgagcggatcgaggaaatcatccggaccaccggcaaagagaacgccaagtacc tgatcgagaagatcaagctgc acgacatgcaggaaggcaagtgcctgtacagcctggaagccatccctctggaagatctgc tgaacaaccccttcaactatgagg tggaccacatcatccccagaagcgtgtccttcgacaacagcttcaacaacaaggtgctcg tgaagcaggaagaagccagcaag aagggcaaccggaccccattccagtacctgagcagcagcgacagcaagatcagctacgaa accttcaagaagcacatcctga atctggccaagggcaagggcagaatcagcaagaccaagaaagagtatctgctggaagaac gggacatcaacaggttctccgt gcagaaagacttcatcaaccggaacctggtggataccagatacgccaccagaggcctgat gaacctgctgcggagctacttca gagtgaacaacctggacgtgaaagtgaagtccatcaatggcggcttcaccagctttctgc ggcggaagtggaagtttaagaaag agcggaacaaggggtacaagcaccacgccgaggacgccctgatcattgccaacgccgatt tcatcttcaaagagtggaagaaa ctggacaaggccaaaaaagtgatggaaaaccagatgttcgaggaaaagcaggccgagagc atgcccgagatcgaaaccgag caggagtacaaagagatcttcatcaccccccaccagatcaagcacattaaggacttcaag gactacaagtacagccaccgggtg gacaagaagcctaatagagagctgattaacgacaccctgtactccacccggaaggacgac aagggcaacaccctgatcgtga acaatctgaacggcctgtacgacaaggacaatgacaagctgaaaaagctgatcaacaaga gccccgaaaagctgctgatgtac caccacgacccccagacctaccagaaactgaagctgattatggaacagtacggcgacgag aagaatcccctgtacaagtacta cgaggaaaccgggaactacctgaccaagtactccaaaaaggacaacggccccgtgatcaa gaagattaagtattacggcaaca aactgaacgcccatctggacatcaccgacgactaccccaacagcagaaacaaggtcgtga agctgtccctgaagccctacaga ttcgacgtgtacctggacaatggcgtgtacaagttcgtgaccgtgaagaatctggatgtg atcaaaaaagaaaactactacgaagt gaatagcaagtgctatgaggaagctaagaagctgaagaagatcagcaaccaggccgagtt tatcgcctccttctacaacaacga tctgatcaagatcaacggcgagctgtatagagtgatcggcgtgaacaacgacctgctgaa ccggatcgaagtgaacatgatcga catcacctaccgcgagtacctggaaaacatgaacgacaagaggccccccaggatcattaa gacaatcgcctccaagacccaga gcattaagaagtacagcacagacattctgggcaacctgtatgaagtgaaatctaagaagc accctcagatcatcaaaaagggca aaaggccggcggccacgaaaaaggccggccaggcaaaaaagaaaaagggatccggacggg ctgacgcattggacgatttt gatctggatatgctgggaagtgacgccctcgatgattttgaccttgacatgcttggttcg gatgcccttgatgactttgacctcgaca tgctcggcagtgacgcccttgatgatttcgacctggacatgctgatttaagaattcacgc gttaagtcgacaatcaacctctggatta caaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgtgg atacgctgctttaatgcctttgtatcatgctat tgcttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctcttta tgaggagttgtggcccgttgtcaggcaacgtg gcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccacct gtcagctcctttccgggactttcgcttt ccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggacagg ggctcggctgttgggcactgacaa ttccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgccac ctggattctgcgcgggacgtccttctgct acgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctctgc ggcctcttccgcgtcttcgccttcgc cctcagacgagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaatga cggccgcaggaacccctagtgatg gagttggccactccctctctgcgcgctcgctcgctcactgaggccgggcgaccaaaggtc gcccgacgcccgggctttgcccg ggcggcctcagtgagcgagcgagcgcgcagctgcctgcaggggcgcctgatgcggtattt tctccttacgcatctgtgcggtatt tcacaccgcatacgtcaaagcaaccatagtacgcgccctgtagcggcgcattaagcgcgg cgggtgtggtggttacgcgcagc gtgaccgctacacttgccagcgccttagcgcccgctcctttcgctttcttcccttccttt ctcgccacgttcgccggctttccccgtca agctctaaatcgggggctccctttagggttccgatttagtgctttacggcacctcgaccc caaaaaacttgatttgggtgatggttca cgtagtgggccatcgccctgatagacggtttttcgccctttgacgttggagtccacgttc tttaatagtggactcttgttccaaactgg aacaacactcaactctatctcgggctattcttttgatttataagggattttgccgatttc ggtctattggttaaaaaatgagctgatttaac aaaaatttaacgcgaattttaacaaaatattaacgtttacaattttatggtgcactctca gtacaatctgctctgatgccgcatagttaa gccagccccgacacccgccaacacccgctgacgcgccctgacgggcttgtctgctcccgg catccgcttacagacaagctgtg accgtctccgggagctgcatgtgtcagaggttttcaccgtcatcaccgaaacgcgcgaga cgaaagggcctcgtgatacgccta tttttataggttaatgtcatgataataatggtttcttagacgtcaggtggcacttttcgg ggaaatgtgcgcggaacccctatttgtttatt tttctaaatacattcaaatatgtatccgctcatgagacaataaccctgataaatgcttca ataatattgaaaaaggaagagtatgagta ttcaacatttccgtgtcgcccttattcccttttttgcggcattttgccttcctgtttttg ctcacccagaaacgctggtgaaagtaaaagat gctgaagatcagttgggtgcacgagtgggttacatcgaactggatctcaacagcggtaag atccttgagagttttcgccccgaag aacgttttccaatgatgagcacttttaaagttctgctatgtggcgcggtattatcccgta ttgacgccgggcaagagcaactcggtc gccgcatacactattctcagaatgacttggttgagtactcaccagtcacagaaaagcatc ttacggatggcatgacagtaagagaa ttatgcagtgctgccataaccatgagtgataacactgcggccaacttacttctgacaacg atcggaggaccgaaggagctaaccg cttttttgcacaacatgggggatcatgtaactcgccttgatcgttgggaaccggagctga atgaagccataccaaacgacgagcg tgacaccacgatgcctgtagcaatggcaacaacgttgcgcaaactattaactggcgaact acttactctagcttcccggcaacaat taatagactggatggaggcggataaagttgcaggaccacttctgcgctcggcccttccgg ctggctggtttattgctgataaatctg gagccggtgagcgtggaagccgcggtatcattgcagcactggggccagatggtaagccct cccgtatcgtagttatctacacga cggggagtcaggcaactatggatgaacgaaatagacagatcgctgagataggtgcctcac tgattaagcattggtaactgtcag accaagtttactcatatatactttagattgatttaaaacttcatttttaatttaaaagga tctaggtgaagatcctttttgataatctcatgac caaaatcccttaacgtgagttttcgttccactgagcgtcagaccccgtagaaaagatcaa aggatcttcttgagatcctttttttctgc gcgtaatctgctgcttgcaaacaaaaaaaccaccgctaccagcggtggtttgtttgccgg atcaagagctaccaactctttttccga aggtaactggcttcagcagagcgcagataccaaatactgttcttctagtgtagccgtagt taggccaccacttcaagaactctgtag caccgcctacatacctcgctctgctaatcctgttaccagtggctgctgccagtggcgata agtcgtgtcttaccgggttggactcaa gacgatagttaccggataaggcgcagcggtcgggctgaacggggggttcgtgcacacagc ccagcttggagcgaacgaccta caccgaactgagatacctacagcgtgagctatgagaaagcgccacgcttcccgaagggag aaaggcggacaggtatccggta agcggcagggtcggaacaggagagcgcacgagggagcttccagggggaaacgcctggtat ctttatagtcctgtcgggtttcg ccacctctgacttgagcgtcgatttttgtgatgctcgtcaggggggcggagcctatggaa aaacgccagcaacgcggcctttttac ggttcctggccttttgctggccttttgctcacatgt pGW045 AAV dSpCas9 Two Vector (SEQ ID NO: 13) cctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtc gggcgacctttggtcgcccgg cctcagtgagcgagcgagcgcgcagagagggagtggccaactccatcactaggggttcct gcggccgcaCGCGTTCT

AGAAAGAgggcctatttcccatgattccttcatatttgcatatacgatacaaggctg ttagagagataattggaattaatttgac tgtaaacacaaagatattagtacaaaatacgtgacgtagaaagtaataatttcttgggta gtttgcagttttaaaattatgttttaaaatg gactatcatatgcttaccgtaacttgaaagtatttcgatttcttggctttatatatcttG TGGAAAGGACGAAACACC ggaagagcgagctcttctgttttagagctaggcc AAC ATGAGGATC ACCC ATGTCTGC AGggcctagca agttaaaataaggctagtccgttatcaacttggccAACATGAGGATCACCCATGTCTGCA Gggccaagt ggcaccgagtcggtgcTTTTTTGGATCCaagcttggcgtaactagatcttgagacaaatg gGACAGCAGAG

ATCC AGTTT GGTT AATT AGCT AGCTGC AAAGATGGAT AAAGTTTT AAAC AGA

GAGGAATCTTTGCAGCTAATGGACCTTCTAGGTCTTGAAAGGAGTGGGAATT

GGCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGA

AGTTGGGGGGAGGGGTCGGCAATTGAACCGGTGCCTAGAGAAGGTGGCGCG

GGGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGT

GGGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCA

ACGGGTTTGCCGCCAGAACACAGGTAAGTGCCGTGTGTGGTTCCCGCGGGCC

TGGCCTCTTTACGGGTTATGGCCCTTGCGTGCCTTGAATTACTTCCACCTGGCT

GC AGT AC GT GATT C TT GATCCC GAGCTTC GGGTT GGA AGT GGGT GGGAG AGT

TCGAGGCCTTGCGCTTAAGGAGCCCCTTCGCCTCGTGCTTGAGTTGAGGCCTG

GCCTGGGCGCTGGGGCCGCCGCGTGCGAATCTGGTGGCACCTTCGCGCCTGT

CTCGCTGCTTTCGATAAGTCTCTAGCCATTTAAAATTTTTGATGACCTGCTGCG

ACGCTTTTTTTCTGGCAAGATAGTCTTGTAAATGCGGGCCAAGATCTGCACAC

TGGTATTTCGGTTTTTGGGGCCGCGGGCGGCGACGGGGCCCGTGCGTCCCAGC

GCACATGTTCGGCGAGGCGGGGCCTGCGAGCGCGGCCACCGAGAATCGGAC

GGGGGTAGTCTCAAGCTGGCCGGCCTGCTCTGGTGCCTGGCCTCGCGCCGCC

GTGTATCGCCCCGCCCTGGGCGGCAAGGCTGGCCCGGTCGGCACCAGTTGCG

T GAGCGGAAAGAT GGCCGCTTCCCGGCCCTGCTGC AGGGAGCTC AAAAT GAA

GGACGCGGCGCTCGGGAGAGCGGGCGGGTGAGTCACCCACACAAAGGAAAA

GGGCCTTTCCGTCCTCAGCCGTCGCTTCATGTGACTCCACGGAGTACCGGGCG

CCGTCCAGGCACCTCGATTAGTTCTCGAGCTTTTGGAGTACGTCGTCTTTAGG

TTGGGGGGAGGGGTTTT AT GCGAT GGAGTTTCCCC AC ACTGAGT GGGT GGAG

ACTGAAGTTAGGCCAGCTTGGCACTTGATGTAATTCTCCTTGGAATTTGCCCT

TTTTGAGTTTGGATCTTGGTTCATTCTCAAGCCTCAGACAGTGGTTCAAAGTTT

TTTTCTTCCATTTCAGGTGTCGTGACGTACGGCCACCATGGCTTCAAACTTTAC

T C AGTTCGT GCTCGT GGAC A ATGGT GGGAC AGGGGAT GT GAC AGT GGCTCCT

TCTAATTTCGCTAATGGGGTGGCAGAGTGGATCAGCTCCAACTCACGGAGCC

AGGCCT AC AAGGT GAC ATGC AGCGT C AGGC AGTCT AGTGCCC AGAAGAGAAA

GT AT AC C ATC A AGGT GGAGGTCCC C A A AGT GGC T AC CC AGAC AGT GGGC GGA

GTCGAACTGCCTGTCGCCGCTTGGAGGTCCTACCTGAACATGGAGCTCACTAT

CCCAATTTTCGCTACCAATTCTGACTGTGAACTCATCGTGAAGGCAATGCAGG

GGCTCCTCAAAGACGGTAATCCTATCCCTTCCGCCATCGCCGCTAACTCAGGT

ATCTACAGCGCTGGAGGAGGTGGAAGCGGAGGAGGAGGAAGCGGAGGAGGA

GGT AGCGGACCT AAGAA AAAGAGGAAGGTGGCGGCCGCTGGATCCCCTT C AG

GGCAGATCAGCAACCAGGCCCTGGCTCTGGCCCCTAGCTCCGCTCCAGTGCT

GGCCCAGACTATGGTGCCCTCTAGTGCTATGGTGCCTCTGGCCCAGCCACCTG

CTCCAGCCCCTGTGCTGACCCCAGGACCACCCCAGTCACTGAGCGCTCCAGTG

CCCAAGTCTACACAGGCCGGCGAGGGGACTCTGAGTGAAGCTCTGCTGCACC

T GC AGTTCGACGCTGAT GAGGACCTGGGAGCTCTGCTGGGGA AC AGC ACCGA

TCCCGGAGTGTTCACAGATCTGGCCTCCGTGGACAACTCTGAGTTTCAGCAGC

TGCTGAATCAGGGCGTGTCCATGTCTCATAGTACAGCCGAACCAATGCTGATG

GAGTACCCCGAAGCCATTACCCGGCTGGTGACCGGCAGCCAGCGGCCCCCCG

ACCCCGCTCCAACTCCCCTGGGAACCAGCGGCCTGCCTAATGGGCTGTCCGG

AGATGAAGACTTCTCAAGCATCGCTGATATGGACTTTAGTGCCCTGCTGTCAC

AGATTTCCTCT AGT GGGC AGGGAGG AGGT GGAAGCGGCTTC AGCGT GGAC AC

CAGTGCCCTGCTGGACCTGTTCAGCCCCTCGGTGACCGTGCCCGACATGAGCC

TGCCTGACCTTGACAGCAGCCTGGCCAGTATCCAAGAGCTCCTGTCTCCCCAG GAGCCCCCCAGGCCTCCCGAGGCAGAGAACAGCAGCCCGGATTCAGGGAAG

CAGCTGGTGCACTACACAGCGCAGCCGCTGTTCCTGCTGGACCCCGGCTCCGT

GGAC ACCGGGAGC A ACGACCTGCCGGT GCTGTTTGAGCTGGGAGAGGGCTCC

TACTTCTCCGAAGGGGACGGCTTCGCCGAGGACCCCACCATCTCCCTGCTGAC

AGGCTCGGAGCCTCCCAAAGCCAAGGACCCCACTGTCTCCTAAGaattcacgcgtta a gtcgacaatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactat gttgctccttttacgctatgtggatacgctg ctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttgt ataaatcctggttgctgtctctttatgaggagttg tggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccact ggttggggcattgccaccacctgtc agctcctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccg cctgccttgcccgctgctggacaggg gctcggctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttcct tggctgctcgcctgtgttgccacctgga ttctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttcctt cccgcggcctgctgccggctctgcgg cctcttccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctcc ccgcgtcgactttaagaccaatgacgt gcggaccgagcggccgcaggaacccctagtgatggagttggccactccctctctgcgcgc tcgctcgctcactgaggccggg cgaccaaaggtcgcccgacgcccgggctttgcccgggcggcctcagtgagcgagcgagcg cgcagctgcctgcaggggcg cctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgcatacgtcaaag caaccatagtacgcgccctgtagcggc gcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgcc ctagcgcccgctcctttcgctttct tcccttcctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctcc ctttagggttccgatttagtgctttacgg cacctcgaccccaaaaaacttgatttgggtgatggttcacgtagtgggccatcgccctga tagacggtttttcgccctttgacgttg gagtccacgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatc tcgggctattcttttgatttataagggattt tgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaatt ttaacaaaatattaacgtttacaattttatg gtgcactctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgcc aacacccgctgacgcgccctgacg ggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcat gtgtcagaggttttcaccgtcatcac cgaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatga taataatggtttcttagacgtcaggtg gcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaa atatgtatccgctcatgagacaataaccct gataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcg cccttattcccttttttgcggcattttgcctt cctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggt gcacgagtgggttacatcgaactgga tctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgag cacttttaaagttctgctatgtggcgcg gtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcag aatgacttggttgagtactcaccagtc acagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataacc atgagtgataacactgcggccaact tacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatggggg atcatgtaactcgccttgatcgttgg gaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagca atggcaacaacgttgcgcaaac tattaactggcgaactacttactctagcttcccggcaacaattaatagactggatggagg cggataaagttgcaggaccacttctgc gctcggcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggt ctcgcggtatcattgcagcactgggg ccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatg gatgaacgaaatagacagatcgct gagataggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatata ctttagattgatttaaaacttcatttttaattt aaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgag ttttcgttccactgagcgtcagaccccg tagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgc aaacaaaaaaaccaccgctaccagcggt ggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcag agcgcagataccaaatactgtccttcta gtgtagccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgct ctgctaatcctgttaccagtggctgct gccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataag gcgcagcggtcgggctgaacggg gggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctaca gcgtgagctatgagaaagcgcc acgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacagga gagcgcacgagggagcttcc agggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcg tcgatttttgtgatgctcgtcaggggg gcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctg gccttttgctcacatgt pRC119 AAV Two Vector dCasl2a/dCpfl (SEQ ID NO: 14) cctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtc gggcgacctttggtcgcccgg cctcagtgagcgagcgagcgcgcagagagggagtggccaactccatcactaggggttcct gcggccgcacgcgttctaggtc ttgaaaggagtgggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccaca gtccccgagaagttggggggag gggtcggcaattgatccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatg tcgtgtactggctccgcctttttcc cgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacgttctttttcgcaa cgggtttgccgccagaacacaggT

GTCGTGACGCGcGTACGgccaccatgacacagttcgagggctttaccaacctgtatc aggtgagcaagacactg cggtttgagctgatcccacagggcaagaccctgaagcacatccaggagcagggcttcatc gaggaggacaaggcccgcaatg atcactacaaggagctgaagcccatcatcgatcggatctacaagacctatgccgaccagt gcctgcagctggtgcagctggatt gggagaacctgagcgccgccatcgactcctatagaaaggagaaaaccgaggagacaagga acgccctgatcgaggagcag gccacatatcgcaatgccatccacgactacttcatcggccggacagacaacctgaccgat gccatcaataagagacacgccga gatctacaagggcctgttcaaggccgagctgtttaatggcaaggtgctgaagcagctggg caccgtgaccacaaccgagcacg agaacgccctgctgcggagcttcgacaagtttacaacctacttctccggcttttatagaa acaggaagaacgtgttcagcgccga ggatatcagcacagccatcccacaccgcatcgtgcaggacaacttccccaagtttaagga gaattgtcacatcttcacacgcctg atcaccgccgtgcccagcctgcgggagcactttgagaacgtgaagaaggccatcggcatc ttcgtgagcacctccatcgagga ggtgttttccttccctttttataaccagctgctgacacagacccagatcgacctgtataa ccagctgctgggaggaatctctcggga ggcaggcaccgagaagatcaagggcctgaacgaggtgctgaatctggccatccagaagaa tgatgagacagcccacatcatc gcctccctgccacacagattcatccccctgtttaagcagatcctgtccgataggaacacc ctgtctttcatcctggaggagtttaag agcgacgaggaagtgatccagtccttctgcaagtacaagacactgctgagaaacgagaac gtgctggagacagccgaggccc tgtttaacgagctgaacagcatcgacctgacacacatcttcatcagccacaagaagctgg agacaatcagcagcgccctgtgcg accactgggatacactgaggaatgccctgtatgagcggagaatctccgagctgacaggca agatcaccaagtctgccaaggag aaggtgcagcgcagcctgaagcacgaggatatcaacctgcaggagatcatctctgccgca ggcaaggagctgagcgaggcct tcaagcagaaaaccagcgagatcctgtcccacgcacacgccgccctggatcagccactgc ctacaaccctgaagaagcagga ggagaaggagatcctgaagtctcagctggacagcctgctgggcctgtaccacctgctgga ctggtttgccgtggatgagtccaa cgaggtggaccccgagttctctgcccggctgaccggcatcaagctggagatggagccttc tctgagcttctacaacaaggccag aaattatgccaccaagaagccctactccgtggagaagttcaagctgaactttcagatgcc tacactggccagaggctgggacgt gaatagagagaagaacaatggcgccatcctgtttgtgaagaacggcctgtactatctggg catcatgccaaagcagaagggca ggtataaggccctgagcttcgagcccacagagaaaaccagcgagggctttgataagatgt actatgactacttccctgatgccgc caagatgatcccaaagtgcagcacccagctgaaggccgtgacagcccactttcagaccca cacaacccccatcctgctgtcca acaatttcatcgagcctctggagatcacaaaggagatctacgacctgaacaatcctgaga aggagccaaagaagtttcagacag cctacgccaagaaaaccggcgaccagaagggctacagagaggccctgtgcaagtggatcg acttcacaagggattttctgtcc aagtataccaagacaacctctatcgatctgtctagcctgcggccatcctctcagtataag gacctgggcgagtactatgccgagct gaatcccctgctgtaccacatcagcttccagagaatcgccgagaaggagatcatggatgc cgtggagacaggcaagctgtacct gttccagatctataacaaggactttgccaagggccaccacggcaagcctaatctgcacac actgtattggaccggcctgttttctc cagagaacctggccaagacaagcatcaagctgaatggccaggccgagctgttctaccgcc ctaagtccaggatgaagaggat ggcacaccggctgggagagaagatgctgaacaagaagctgaaggatcagaaaaccccaat ccccgacaccctgtaccagga gctgtacgactatgtgaatcacagactgtcccacgacctgtctgatgaggccagggccct gctgcccaacgtgatcaccaagga ggtgtctcacgagatcatcaaggataggcgctttaccagcgacaagttctttttccacgt gcctatcacactgaactatcaggccgc caattccccatctaagttcaaccagagggtgaatgcctacctgaaggagcaccccgagac acctatcatcggcatcgcccgggg cgagagaaacctgatctatatcacagtgatcgactccaccggcaagatcctggagcagcg gagcctgaacaccatccagcagtt tgattaccagaagaagctggacaacagggagaaggagagggtggcagcaaggcaggcctg gtctgtggtgggcacaatcaa ggatctgaagcagggctatctgagccaggtcatccacgagatcgtggacctgatgatcca ctaccaggccgtggtggtgctgga gaacctgaatttcggctttaagagcaagaggaccggcatcgccgagaaggccgtgtacca gcagttcgagaagatgctgatcg ataagctgaattgcctggtgctgaaggactatccagcagagaaagtgggaggcgtgctga acccataccagctgacagaccag ttcacctcctttgccaagatgggcacccagtctggcttcctgttttacgtgcctgcccca tatacatctaagatcgatcccctgaccg gcttcgtggaccccttcgtgtggaaaaccatcaagaatcacgagagccgcaagcacttcc tggagggcttcgactttctgcacta cgacgtgaaaaccggcgacttcatcctgcactttaagatgaacagaaatctgtccttcca gaggggcctgcccggctttatgcctg catgggatatcgtgttcgagaagaacgagacacagtttgacgccaagggcacccctttca tcgccggcaagagaatcgtgcca gtgatcgagaatcacagattcaccggcagataccgggacctgtatcctgccaacgagctg atcgccctgctggaggagaaggg catcgtgttcagggatggctccaacatcctgccaaagctgctggagaatgacgattctca cgccatcgacacgatggtggccctg atccgcagcgtgctgcagatgcggaactccaatgccgccacaggcgaggactatatcaac agccccgtgcgcgatctgaatgg cgtgtgcttcgactcccggtttcagaacccagagtggccaatggacgccgatgccaatgg cgcctaccacatcgccctgaagg gccagctgctgctgaatcacctgaaggagagcaaggatctgaagctgcagaacggcatct ccaatcaggactggctggcctac atccaggagctgcgcaacaaaaggccggcggccacgaaaaaggccggccaggcaaaaaag aaaaagGGCAGTGG

T AGT GGAGA AGAGC T GTT GT C A A A A A ATT ATC ACTTGGAGA AT GAGGT AGCC CGCCTTAAGAAAGGCAGCGGATCAGGCGAGGAACTTCTGAGCAAGAATTACC

ATCTCGAAAACGAAGTGGCACGCTTGAAGAAGGGTTCAGGTTCCGGGGAGGA

GCTCCTCTCCAAGAACTACCACCTGGAGAACGAAGTCGCTCGCCTGAAGAAAt gaAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGcggac cgagcggccgcaggaacccctagtgatggagttggccactccctctctgcgcgctcgctc gctcactgaggccgggcgacca aaggtcgcccgacgcccgggctttgcccgggcggcctcagtgagcgagcgagcgcgcagc tgcctgcaggggcgcctgat gcggtattttctccttacgcatctgtgcggtatttcacaccgcatacgtcaaagcaacca tagtacgcgccctgtagcggcgcatta agcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcg cccgctcctttcgctttcttcccttc ctttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttagg gttccgatttagtgctttacggcacctc gaccccaaaaaacttgatttgggtgatggttcacgtagtgggccatcgccctgatagacg gtttttcgccctttgacgttggagtcc acgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcgggc tattcttttgatttataagggattttgccga tttcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaaca aaatattaacgtttacaattttatggtgcact ctcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacaccc gctgacgcgccctgacgggcttgt ctgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcag aggttttcaccgtcatcaccgaaac gcgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataa tggtttcttagacgtcaggtggcactttt cggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtat ccgctcatgagacaataaccctgataaat gcttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttat tcccttttttgcggcattttgccttcctgtttt tgctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagt gggttacatcgaactggatctcaac agcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcactttt aaagttctgctatgtggcgcggtattatc ccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgactt ggttgagtactcaccagtcacagaa aagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagt gataacactgcggccaacttacttct gacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgt aactcgccttgatcgttgggaacc ggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggc aacaacgttgcgcaaactattaa ctggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggata aagttgcaggaccacttctgcgctcg gcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgc ggtatcattgcagcactggggccag atggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatg aacgaaatagacagatcgctgagat aggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatacttta gattgatttaaaacttcatttttaatttaaaa ggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagtttt cgttccactgagcgtcagaccccgtaga aaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaac aaaaaaaccaccgctaccagcggtggtt tgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcg cagataccaaatactgtccttctagtgt agccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgc taatcctgttaccagtggctgctgcca gtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgc agcggtcgggctgaacggggggt tcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgt gagctatgagaaagcgccacgct tcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcg cacgagggagcttccaggg ggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcga tttttgtgatgctcgtcaggggggcgg agcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcct tttgctcacatgt pRC120 AAV Two Vector dCasl2a/dCpfl (SEQ ID NO: 15) cctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtc gggcgacctttggtcgcccgg cctcagtgagcgagcgagcgcgcagagagggagtggccaactccatcactaggggttcct gcggccgcacgcgttctaggtc ttgaaaggagtgggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccaca gtccccgagaagttggggggag gggtcggcaattgatccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatg tcgtgtactggctccgcctttttcc cgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacgttctttttcgcaa cgggtttgccgccagaacacaggT

GTCGTGACGCGcGTACGgccaccatgtcaaagctggagaaattcaccaactgttata gcctgtctaagaccctgc gcttcaaggcaatcccagtgggcaagacacaagagaacattgacaacaaacggctcctgg tggaggatgagaagagggctga agattacaagggcgttaagaagctgctggataggtactatctgtcattcatcaacgatgt cctccacagtatcaagctgaagaatct gaacaattacatttctctgttccggaagaagacacggaccgagaaggagaacaaagagct ggagaatctggagatcaacctga ggaaagaaatagctaaggctttcaaagggaacgagggttacaagtccctgttcaagaaag acattatcgagactattctgcctga gttcctggacgataaagatgagatcgccctcgtcaattccttcaatgggtttaccacagc ctttaccggcttcttcgacaatagaga gaatatgttctctgaagaggccaaatccactagcatcgcctttcgctgcataaacgagaa cctgactaggtacatcagcaatatgg acatctttgagaaagtcgatgccatattcgacaaacatgaggtgcaggagattaaggaga agatcctgaactcagattacgatgtc gaagatttcttcgagggagagttcttcaacttcgtgctcacacaagagggcattgatgtg tacaatgcaatcattggagggttcgtg acagagagtggcgagaagataaagggcctgaacgagtatatcaacctctacaaccagaaa accaagcagaaactgcctaagtt caagccactgtacaaacaagtgctctcagatagggaaagcctgagcttctacggtgaagg gtatacatcagatgaagaagtgct cgaagtgttccgcaacaccctcaataagaacagtgaaatcttctcttcaatcaagaagct ggagaaactgttcaagaatttcgatga gtactcctctgccggaatctttgtgaagaatggccctgcaatatccactattagcaaaga catctttggcgagtggaacgttatcag ggataagtggaatgccgagtacgatgatattcatctcaagaagaaagccgtggttacaga gaaatacgaggatgatagacgcaa gagctttaagaagattggtagcttctctctcgaacagctgcaggagtacgccgacgctga cctgtcagtcgtggagaaactcaag gagatcataatccagaaggtggatgaaatctacaaagtgtatggaagctctgagaaactc ttcgatgcagactttgttctggagaa gagtctgaagaagaacgacgcagtggttgctatcatgaaggacctgctggattctgttaa gtctttcgagaattacattaaggcatt ctttggtgaagggaaggagacaaatagggacgagagcttctatggcgactttgttctggc ctacgacatcctcctcaaggttgacc acatctatgacgctatacggaattacgttacccagaagccctatagcaaagacaagttca agctgtatttccagaatccacagtttat gggtgggtgggataaagacaaagaaacagattacagggccactatcctgcggtacggcag caaatactatctggctatcatgga taagaagtacgccaaatgcctccagaagatcgacaaggacgacgtgaacggtaactacga gaagatcaattacaagctcctgc caggacctaacaagatgctgcccaaggtgttcttctccaagaaatggatggcctactata acccaagcgaggacattcagaagat atacaagaatgggacattcaagaagggcgatatgttcaacctcaacgactgccacaagct gattgatttcttcaaggatagcatttc tcgctatcccaagtggtctaatgcatacgatttcaacttcagcgagactgagaagtacaa agacatcgctggcttctaccgggagg tggaagagcaaggctataaggtgtcattcgaatccgcttctaagaaggaagtggataagc tcgtggaagagggtaagctgtacat gttccagatatacaacaaagacttcagcgataagagccacggcactccaaacctccatac tatgtatttcaagctgctgtttgacga gaacaaccacggacagattaggctgtcaggaggcgcagaactcttcatgcgcagagcttc actgaagaaggaggaactcgttg tccacccagccaatagccctatagccaataagaatccagacaatcctaagaaaaccacta ctctgtcttacgatgtgtataaggat aagagattctctgaagatcagtacgaactgcacatacccattgccattaacaagtgccct aagaacatcttcaagattaacacaga ggttagagtgctcctgaaacacgacgataacccttatgttataggcattgctcgcggaga gagaaacctgctgtacatcgtcgtgg tggacggcaaaggcaacatcgtggaacagtacagtctcaatgaaatcattaacaatttca acggaatccgcattaagaccgacta ccattctctcctcgacaagaaggagaaagaaaggttcgaagcaagacagaattggacaag tatagagaatatcaaagaactgaa ggctgggtacatctctcaggttgtgcacaagatatgtgagctggtggagaagtacgacgc tgttatcgccctcgcggacctgaat agcggcttcaagaactccagggtgaaggtggagaagcaggtgtatcagaagttcgagaag atgctgatcgacaagctcaactat atggtggacaagaaatccaatccttgcgctactggtggagccctgaagggctatcaaatc accaataagttcgaatctttcaagtct atgagcacccagaatggcttcatcttctacatacccgcatggctgacatccaagattgat ccctctaccggatttgttaatctgctca agactaagtacacctctattgctgactcaaagaagttcatatcatcatttgaccgcatca tgtacgtgccagaagaggacctgttcg agtttgccctggattacaagaatttctctcggactgacgccgactacatcaagaagtgga agctctactcttatggtaatcggattcg catattccgcaatcccaagaagaataacgtgttcgattgggaggaagtttgcctcaccag cgcttacaaggagctgttcaataagt atgggattaactaccagcagggcgacataagagccctgctgtgcgaacaatctgataagg cattctattcctctttcatggcactga tgtcactgatgctgcaaatgcgcaattccatcaccggaagaacagacgtggcctttctga tctctcctgtcaagaactcagatggc atcttctacgattcccgcaactatgaagcacaggagaatgctatcctgcctaagaatgcc gatgcaaatggagcctataacatcgc cagaaaggtcctctgggccataggacaattcaagaaagctgaagatgagaagctggacaa ggtgaagatcgccatttcaaaca aagagtggctcgaatatgctcagacctcagtgaagcataaaaggccggcggccacgaaaa aggccggccaggcaaaaaaga aaaagGGC AGCGGT AGT GGGGAGGAGCTTCTTTCT AAAAATT AT C ATCTTGAAA

AT GAAGT GGC ACGACTC AAGAAAGGCTCTGGGTCCGGT GAGGAACTCCTGTC

CAAGAACTATCATCTTGAAAACGAAGTCGCCCGCCTTAAAAAAGGCTCAGGC

TCAGGTGAAGAATTGTTGAGCAAAAACTATCACCTTGAGAATGAGGTTGCCC

GGCTCAAAAAAGGGTCTGGCAGCGGCGAGGAACTCCTTTCAAAGAACTACCA

TCTCGAAAATGAAGTTGCTCGGTT GAAGAAGGGTTC AGGATCTGGCGAGGAG

CTTCTGTCCAAGAACTACCATTTGGAGAACGAGGTCGCAAGACTTAAAAAAtg aAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGcggacc gagcggccgcaggaacccctagtgatggagttggccactccctctctgcgcgctcgctcg ctcactgaggccgggcgaccaa aggtcgcccgacgcccgggctttgcccgggcggcctcagtgagcgagcgagcgcgcagct gcctgcaggggcgcctgatg cggtattttctccttacgcatctgtgcggtatttcacaccgcatacgtcaaagcaaccat agtacgcgccctgtagcggcgcattaa gcgcggcgggtgtggtggttacgcgcagcgtgaccgctacacttgccagcgccctagcgc ccgctcctttcgctttcttcccttcc tttctcgccacgttcgccggctttccccgtcaagctctaaatcgggggctccctttaggg ttccgatttagtgctttacggcacctcg accccaaaaaacttgatttgggtgatggttcacgtagtgggccatcgccctgatagacgg tttttcgccctttgacgttggagtcca cgttctttaatagtggactcttgttccaaactggaacaacactcaaccctatctcgggct attcttttgatttataagggattttgccgatt tcggcctattggttaaaaaatgagctgatttaacaaaaatttaacgcgaattttaacaaa atattaacgtttacaattttatggtgcactc tcagtacaatctgctctgatgccgcatagttaagccagccccgacacccgccaacacccg ctgacgcgccctgacgggcttgtc tgctcccggcatccgcttacagacaagctgtgaccgtctccgggagctgcatgtgtcaga ggttttcaccgtcatcaccgaaacg cgcgagacgaaagggcctcgtgatacgcctatttttataggttaatgtcatgataataat ggtttcttagacgtcaggtggcacttttc ggggaaatgtgcgcggaacccctatttgtttatttttctaaatacattcaaatatgtatc cgctcatgagacaataaccctgataaatg cttcaataatattgaaaaaggaagagtatgagtattcaacatttccgtgtcgcccttatt cccttttttgcggcattttgccttcctgttttt gctcacccagaaacgctggtgaaagtaaaagatgctgaagatcagttgggtgcacgagtg ggttacatcgaactggatctcaac agcggtaagatccttgagagttttcgccccgaagaacgttttccaatgatgagcactttt aaagttctgctatgtggcgcggtattatc ccgtattgacgccgggcaagagcaactcggtcgccgcatacactattctcagaatgactt ggttgagtactcaccagtcacagaa aagcatcttacggatggcatgacagtaagagaattatgcagtgctgccataaccatgagt gataacactgcggccaacttacttct gacaacgatcggaggaccgaaggagctaaccgcttttttgcacaacatgggggatcatgt aactcgccttgatcgttgggaacc ggagctgaatgaagccataccaaacgacgagcgtgacaccacgatgcctgtagcaatggc aacaacgttgcgcaaactattaa ctggcgaactacttactctagcttcccggcaacaattaatagactggatggaggcggata aagttgcaggaccacttctgcgctcg gcccttccggctggctggtttattgctgataaatctggagccggtgagcgtgggtctcgc ggtatcattgcagcactggggccag atggtaagccctcccgtatcgtagttatctacacgacggggagtcaggcaactatggatg aacgaaatagacagatcgctgagat aggtgcctcactgattaagcattggtaactgtcagaccaagtttactcatatatacttta gattgatttaaaacttcatttttaatttaaaa ggatctaggtgaagatcctttttgataatctcatgaccaaaatcccttaacgtgagtttt cgttccactgagcgtcagaccccgtaga aaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatctgctgcttgcaaac aaaaaaaccaccgctaccagcggtggtt tgtttgccggatcaagagctaccaactctttttccgaaggtaactggcttcagcagagcg cagataccaaatactgtccttctagtgt agccgtagttaggccaccacttcaagaactctgtagcaccgcctacatacctcgctctgc taatcctgttaccagtggctgctgcca gtggcgataagtcgtgtcttaccgggttggactcaagacgatagttaccggataaggcgc agcggtcgggctgaacggggggt tcgtgcacacagcccagcttggagcgaacgacctacaccgaactgagatacctacagcgt gagctatgagaaagcgccacgct tcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcggaacaggagagcg cacgagggagcttccaggg ggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctgacttgagcgtcga tttttgtgatgctcgtcaggggggcgg agcctatggaaaaacgccagcaacgcggcctttttacggttcctggccttttgctggcct tttgctcacatgt pRC121b AAV Two Vector dCasl2a/dCpfl (SEQ ID NO: 16)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgGGCCCAGCCGCCAAACGAGTCAAGCTGGATGGCCCCGATATCGTCATGAC

TCAATCACCCTCATCACTGTCAGCCAGCGTCGGTGACCGGGTAACAATTACTT

GTAGATCATCAACTGGGGCCGTCACTACCTCCAACTATGCCAGTTGGGTTCAA

GAGAAACCCGGCAAGCTCTTTAAGGGACTGATCGGTGGGACCAATAATCGGG

CCCCCGGTGTGCCTTCTCGCTTTAGCGGGAGTTTGATAGGCGATAAAGCCACA

TTGACAATATCATCTCTTCAACCCGAGGACTTCGCTACCTATTTTTGTGCTTTG

T GGT AT AGT A ACC ATT GGGT ATTTGGC C A AGGA ACT A AGGT AGAGTT GA AGC

GGGGGGGCGGCGGCTCTGGTGGAGGAGGTAGTGGGGGCGGTGGCTCTTCCGG AGGGGGTTCCGAAGTAAAGCTTTTGGAGAGCGGCGGCGGCCTGGTCCAGCCT

GGAGGATCTCTCAAGCTTAGCTGCGCCGTCAGCGGCTTTTCCCTCACTGATTA

T GGTGT AAATT GGGT GAGGC AGGCTCC AGGC AGGGGGTT GGAGT GGAT AGGG

GTGATATGGGGGGATGGCATCACAGATTATAATTCTGCACTTAAGGATAGGT

TTATTATCAGTAAGGACAACGGTAAGAACACCGTTTATCTTCAGATGTCTAAG

GTGCGGTCCGATGATACTGCCCTCTATTATTGTGTAACCGGCCTTTTCGATTAC

TGGGGCCAAGGGACTTTGGTAACCGTGTCATCATATCCCTATGACGTCCCTGA

C T AC GC AGGGGG AGG AGGGGG AT C C GGT GGC GGT GGT AGT GG AGG AGGT GG

ATCTGGAGGAGGGGGAAGCggtaccCC AGCTGCGAAACGAGTT AAATTGGATGG

T GGT GGC GGGT C CTC AGGTTT GCC C A AT GGATTGGAT GGT GAT GA AGAC TTC T

CAGACATCGCAGATATGGACTTTAGTGCCTTGCTGAGCCAAATCTCATCTGGC

TCATCAGGTCTTCCCAACGGATTGGATGGCGATGAGGACTTTTCCGACATCGC

AGACATGGACTTTTCCGCTCTGTTGAGCCAGATAAGTTCTGGCTCCTCCGGAC

TCCCTAATGGCTTGGATGGAGATGAGGATTTTAGCGATATAGCAGACATGGA

TTTTAGCGCATTGCTCTCACAAATCTCCAGTGGCGGGGGTGGTAGTGGATTTT

CAGTGGATACTTCCGCACTTCTCGATTTGTTTTCTCCCAGTGTCACCGTCCCAG

ACATGAGTCTTCCTGACCTCGACTCTTCTCTCGCCAGTATTCAGGAACTGCTC

AGTCCACAGGAGCCTCCTCGCCCTCCTGAAGCAGAGAATAGTTCCCCAGATT

CAGGAAAGCAGCTGGTGCACTACACAGCACAGCCACTGTTCCTCTTGGACCC

TGGGAGTGTAGACACAGGCTCAAACGATCTCCCCGTTCTTTTTGAACTTGGCG

AAGGTTCCTATTTTAGCGAGGGGGACGGATTCGCCGAGGACCCTACTATCAG

TCTCCTTACAGGTAGCGAGCCACCCAAAGCTAAAGACCCCACCGTGAGCTAG

TGActcgagGATTCGTCAGTAGGGTTGTAAAGGTTTTTCTTTTCCTGAGAAAACA

ACCTTTTGTTTTCTCAGGTTTTGCTTTTTGGCCTTTCCCTAGCTTTAAAAAAAA

AAAAGCAAAACTCACCGAGGCAGTTCCATAGGATGGCtagcAAGATCCTGGTA

TT GGTCTGCGAgaagagcgagctcttcaatcaacctctggattacaaaatttgtgaaagatt gactggtattcttaactatg ttgctccttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgctt cccgtatggctttcattttctcctccttgtataaa tcctggttgctgtctctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtg tgcactgtgtttgctgacgcaaccccc actggttggggcattgccaccacctgtcagctcctttccgggactttcgctttccccctc cctattgccacggcggaactcatcgcc gcctgccttgcccgctgctggacaggggctcggctgttgggcactgacaattccgtggtg ttgtcggggaaatcatcgtcctttcc ttggctgctcgcctgtgttgccacctggattctgcgcgggacgtccttctgctacgtccc ttcggccctcaatccagcggaccttcc ttcccgcggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagac gagtcggatctccctttgggccgcct ccccgcgtcgactttaagaccaatCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACT CC

CTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGA

CGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCC

TGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCA

CACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAG

CGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCC

TTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGC

TTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGC

TTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTG

GGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTC

TTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGG

CTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAA

TGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTA

CAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGC

CAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGC

TCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGT

CAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGA

TACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCA GGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTA

AATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTC

AATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTT

ATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTG

GTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCG

AACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACG

TTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCC

GTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAA

TGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATG

ACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGG

CCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTG

CACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGA

ATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGC

AACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGC

AACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG

CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGC

GTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCG

T ATC GT AGT TATCTACAC G AC GGGG AGT C AGGC A AC TAT GG AT G A AC G A A AT

AGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAG

ACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTA

AAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAA

CGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGAT

CTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA

CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTT

TCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAG

TGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATAC

CTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTG

TCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCG

GGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACA

CCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGA

AGGGAGA A AGGC GGAC AGGT ATCCGGT A AGC GGC AGGGTCGGA AC AGGAGA

GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTC

GGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGG

GCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCC

TTTTGCTGGCCTTTTGCTCACATGT pRC124 AAV Two Vector dCasl2a/dCpfl (SEQ ID NO: 17)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgCCCGCTGCGAAACGAGTTAAATTGGATggatcctcaaagctggagaaattcaccaac tgttatagc ctgtctaagaccctgcgcttcaaggcaatcccagtgggcaagacacaagagaacattgac aacaaacggctcctggtggagga tgagaagagggctgaagattacaagggcgttaagaagctgctggataggtactatctgtc attcatcaacgatgtcctccacagta tcaagctgaagaatctgaacaattacatttctctgttccggaagaagacacggaccgaga aggagaacaaagagctggagaatc tggagatcaacctgaggaaagaaatagctaaggctttcaaagggaacgagggttacaagt ccctgttcaagaaagacattatcg agactattctgcctgagttcctggacgataaagatgagatcgccctcgtcaattccttca atgggtttaccacagcctttaccggctt cttcgacaatagagagaatatgttctctgaagaggccaaatccactagcatcgcctttcg ctgcataaacgagaacctgactaggt acatcagcaatatggacatctttgagaaagtcgatgccatattcgacaaacatgaggtgc aggagattaaggagaagatcctgaa ctcagattacgatgtcgaagatttcttcgagggagagttcttcaacttcgtgctcacaca agagggcattgatgtgtacaatgcaatc attggagggttcgtgacagagagtggcgagaagataaagggcctgaacgagtatatcaac ctctacaaccagaaaaccaagca gaaactgcctaagttcaagccactgtacaaacaagtgctctcagatagggaaagcctgag cttctacggtgaagggtatacatca gatgaagaagtgctcgaagtgttccgcaacaccctcaataagaacagtgaaatcttctct tcaatcaagaagctggagaaactgtt caagaatttcgatgagtactcctctgccggaatctttgtgaagaatggccctgcaatatc cactattagcaaagacatctttggcga gtggaacgttatcagggataagtggaatgccgagtacgatgatattcatctcaagaagaa agccgtggttacagagaaatacga ggatgatagacgcaagagctttaagaagattggtagcttctctctcgaacagctgcagga gtacgccgacgctgacctgtcagtc gtggagaaactcaaggagatcataatccagaaggtggatgaaatctacaaagtgtatgga agctctgagaaactcttcgatgcag actttgttctggagaagagtctgaagaagaacgacgcagtggttgctatcatgaaggacc tgctggattctgttaagtctttcgaga attacattaaggcattctttggtgaagggaaggagacaaatagggacgagagcttctatg gcgactttgttctggcctacgacatc ctcctcaaggttgaccacatctatgacgctatacggaattacgttacccagaagccctat agcaaagacaagttcaagctgtatttc cagaatccacagtttatgggtgggtgggataaagacaaagaaacagattacagggccact atcctgcggtacggcagcaaatac tatctggctatcatggataagaagtacgccaaatgcctccagaagatcgacaaggacgac gtgggtaccagtCCAGCTG

CGAAACGAGTTAAATTGGATGGTGGTGGCGGGTCCTCAGGTTTGCCCAATGG

ATTGGATGGTGATGAAGACTTCTCAGACATCGCAGATATGGACTTTAGTGCCT

TGCTGAGCCAAATCTCATCTGGCTCATCAGGTCTTCCCAACGGATTGGATGGC

GATGAGGACTTTTCCGACATCGCAGACATGGACTTTTCCGCTCTGTTGAGCCA

GATAAGTTCTGGCTCCTCCGGACTCCCTAATGGCTTGGATGGAGATGAGGATT

TTAGCGATATAGCAGACATGGATTTTAGCGCATTGCTCTCACAAATCTCCAGT

GGCGGGGGTGGTAGTGGATTTTCAGTGGATACTTCCGCACTTCTCGATTTGTT

TTCTCCCAGTGTCACCGTCCCAGACATGAGTCTTCCTGACCTCGACTCTTCTCT

CGCCAGTATTCAGGAACTGCTCAGTCCACAGGAGCCTCCTCGCCCTCCTGAAG

CAGAGAATAGTTCCCCAGATTCAGGAAAGCAGCTGGTGCACTACACAGCACA

GCCACTGTTCCTCTTGGACCCTGGGAGTGTAGACACAGGCTCAAACGATCTCC

CCGTTCTTTTTGAACTTGGCGAAGGTTCCTATTTTAGCGAGGGGGACGGATTC

GCCGAGGACCCTACTATCAGTCTCCTTACAGGTAGCGAGCCACCCAAAGCTA

AAGACCCCACCGTGAGCTAGTGActcgagGATTCGTCAGTAGGGTTGTAAAGGT

TTTTCTTTTCCTGAGAAAACAACCTTTTGTTTTCTCAGGTTTTGCTTTTTGGCCT

TTCCCTAGCTTTAAAAAAAAAAAAGCAAAACTCACCGAGGCAGTTCCATAGG

AT GGCtagc A AG AT C CTGGT ATTGGT C T GC GAgaagagcgagctcttcaatcaacctctggattacaa aatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgtggata cgctgctttaatgcctttgtatcatgctattgct tcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctttatgag gagttgtggcccgttgtcaggcaacgtggcg tggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccacctgtc agctcctttccgggactttcgctttccc cctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggacaggggc tcggctgttgggcactgacaattcc gtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgccacctgg attctgcgcgggacgtccttctgctacg tcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctctgcggc ctcttccgcgtcttcgccttcgccctc agacgagtcggatctccctttgggccgcctccccgcCGGCCGCAGGAACCCCTAGTGATG GAGTTG

GCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGT

CGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC

AGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGG

TATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCG

CATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGC CAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTT

CGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGAT

TTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCA

CGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTC

CACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTA

TCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGT

TAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATT

AACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCAT

AGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGC

TTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCT

GCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGG

CCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTA

GACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTAT

TTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAA

ATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGT

CGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGA

A AC GC T GGT G A A AGT A A A AG AT GC T G A AG AT C AGT T GGGT GC AC G AGT GGGT

TACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGA

AGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTAT

TATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCT

CAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATG

GCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACAC

TGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCT

TTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGA

GCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCA

ATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTC

CCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTT

CTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGG

TGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCC

TCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAAC

GAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACT

GTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTA

ATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCC

CTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAA

GGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAA

AAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCT

TTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTC

TAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACA

TACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTC

GTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG

TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCT

ACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCC

CGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGG

AGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCT

GTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGG

GGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTG

GCCTTTTGCTGGCCTTTTGCTCACATGT pRC126 AAV Two Vector dCasl2a/dCpfl (SEQ ID NO: 18) CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGCCCGCTGCGAAACGAGTTAAATTGGATGGTGGTGGCGGGTCCTCAGGTT

TGCCCAATGGATTGGATGGTGATGAAGACTTCTCAGACATCGCAGATATGGA

CTTTAGTGCCTTGCTGAGCCAAATCTCATCTGGCTCATCAGGTCTTCCCAACG

GATTGGATGGCGATGAGGACTTTTCCGACATCGCAGACATGGACTTTTCCGCT

CTGTTGAGCCAGATAAGTTCTGGCTCCTCCGGACTCCCTAATGGCTTGGATGG

AGATGAGGATTTTAGCGATATAGCAGACATGGATTTTAGCGCATTGCTCTCAC

AAATCTCCAGTGGCGGGGGTGGTAGTGGATTTTCAGTGGATACTTCCGCACTT

CTCGATTTGTTTTCTCCCAGTGTCACCGTCCCAGACATGAGTCTTCCTGACCTC

GACTCTTCTCTCGCCAGTATTCAGGAACTGCTCAGTCCACAGGAGCCTCCTCG

CCCTCCTGAAGCAGAGAATAGTTCCCCAGATTCAGGAAAGCAGCTGGTGCAC

TACACAGCACAGCCACTGTTCCTCTTGGACCCTGGGAGTGTAGACACAGGCTC

AAACGATCTCCCCGTTCTTTTTGAACTTGGCGAAGGTTCCTATTTTAGCGAGG

GGGACGGATTCGCCGAGGACCCTACTATCAGTCTCCTTACAGGTAGCGAGCC

ACCCAAAGCTAAAGACCCCACCGTGAGCGGTGGTGGCGGGTCCGGTaacggtaact acgagaagatcaattacaagctcctgccaggacctaacaagatgctgcccaaggtgttct tctccaagaaatggatggcctactat aacccaagcgaggacattcagaagatatacaagaatgggacattcaagaagggcgatatg ttcaacctcaacgactgccacaa gctgattgatttcttcaaggatagcatttctcgctatcccaagtggtctaatgcatacga tttcaacttcagcgagactgagaagtaca aagacatcgctggcttctaccgggaggtggaagagcaaggctataaggtgtcattcgaat ccgcttctaagaaggaagtggata agctcgtggaagagggtaagctgtacatgttccagatatacaacaaagacttcagcgata agagccacggcactccaaacctcc atactatgtatttcaagctgctgtttgacgagaacaaccacggacagattaggctgtcag gaggcgcagaactcttcatgcgcaga gcttcactgaagaaggaggaactcgttgtccacccagccaatagccctatagccaataag aatccagacaatcctaagaaaacc actactctgtcttacgatgtgtataaggataagagattctctgaagatcagtacgaactg cacatacccattgccattaacaagtgcc ctaagaacatcttcaagattaacacagaggttagagtgctcctgaaacacgacgataacc cttatgttataggcattgctcgcgga gagagaaacctgctgtacatcgtcgtggtggacggcaaaggcaacatcgtggaacagtac agtctcaatgaaatcattaacaatt tcaacggaatccgcattaagaccgactaccattctctcctcgacaagaaggagaaagaaa ggttcgaagcaagacagaattgga caagtatagagaatatcaaagaactgaaggctgggtacatctctcaggttgtgcacaaga tatgtgagctggtggagaagtacga cgctgttatcgccctcgcggacctgaatagcggcttcaagaactccagggtgaaggtgga gaagcaggtgtatcagaagttcga gaagatgctgatcgacaagctcaactatatggtggacaagaaatccaatccttgcgctac tggtggagccctgaagggctatcaa atcaccaataagttcgaatctttcaagtctatgagcacccagaatggcttcatcttctac atacccgcatggctgacatccaagattg atccctctaccggatttgttaatctgctcaagactaagtacacctctattgctgactcaa agaagttcatatcatcatttgaccgcatca tgtacgtgccagaagaggacctgttcgagtttgccctggattacaagaatttctctcgga ctgacgccgactacatcaagaagtgg aagctctactcttatggtaatcggattcgcatattccgcaatcccaagaagaataacgtg ttcgattgggaggaagtttgcctcacc agcgcttacaaggagctgttcaataagtatgggattaactaccagcagggcgacataaga gccctgctgtgcgaacaatctgata aggcattctattcctctttcatggcactgatgtcactgatgctgcaaatgcgcaattcca tcaccggaagaacagacgtggcctttct gatctctcctgtcaagaactcagatggcatcttctacgattcccgcaactatgaagcaca ggagaatgctatcctgcctaagaatgc cgatgcaaatggagcctataacatcgccagaaaggtcctctgggccataggacaattcaa gaaagctgaagatgagaagctgg acaaggtgaagatcgccatttcaaacaaagagtggctcgaatatgctcagacctcagtga agcatggtaccCCCGCTGC GAAACGAGTTAAATTGGATGGTGGTGGCGGGTCCTCAGGTTTGCCCAATGGA

TTGGATGGTGATGAAGACTTCTCAGACATCGCAGATATGGACTTTAGTGCCTT

GCTGAGCCAAATCTCATCTGGCTCATCAGGTCTTCCCAACGGATTGGATGGCG

ATGAGGACTTTTCCGACATCGCAGACATGGACTTTTCCGCTCTGTTGAGCCAG

ATAAGTTCTGGCTCCTCCGGACTCCCTAATGGCTTGGATGGAGATGAGGATTT

TAGCGATATAGCAGACATGGATTTTAGCGCATTGCTCTCACAAATCTCCAGTG

GCGGGGGTGGTAGTGGATTTTCAGTGGATACTTCCGCACTTCTCGATTTGTTT

TCTCCCAGTGTCACCGTCCCAGACATGAGTCTTCCTGACCTCGACTCTTCTCTC

GCCAGTATTCAGGAACTGCTCAGTCCACAGGAGCCTCCTCGCCCTCCTGAAGC

AGAGAATAGTTCCCCAGATTCAGGAAAGCAGCTGGTGCACTACACAGCACAG

CCACTGTTCCTCTTGGACCCTGGGAGTGTAGACACAGGCTCAAACGATCTCCC

CGTTCTTTTTGAACTTGGCGAAGGTTCCTATTTTAGCGAGGGGGACGGATTCG

CCGAGGACCCTACTATCAGTCTCCTTACAGGTAGCGAGCCACCCAAAGCTAA

AGACCCCACCGTGAGCTAGTGActcgagAATAAAAGATCTTTATTTTCATTAGAT

CTGTGTGTTGGTTTTTTGTGTCGGCCGCAGGAACCCCTAGTGATGGAGTTGGC

CACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCG

CCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAG

CTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTA

TTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCA

TTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCA

GCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCG

CCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTT

AGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACG

TAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCA

CGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATC

TCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTA

AAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAA

CGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAG

TTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTT

GTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGC

ATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCC

TCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGA

CGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTT

TTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAAT

GCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCG

CCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAA

C GC T GGT G A A AGT A A A AG AT GC T G A AG AT C AGTT GGGT GC AC G AGT GGGT T A

CATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAA

GAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATT

ATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTC

AGAAT GACTTGGTT GAGT ACTC ACC AGT C AC AGA AAAGC ATCTT ACGGAT GG

CAT GAC AGT AAGAGAATT AT GC AGT GCTGCC AT AACC AT GAGT GAT AAC ACT

GCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTT

TTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAG

CTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAA

TGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCC

CGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTC

TGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGT

GAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCT

CCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACG AAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTG

TCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAA

TTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCC

TTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAA

GGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAA

AAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCT

TTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTC

TAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACA

TACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTC

GTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG

TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCT

ACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCC

CGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGG

AGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCT

GTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGG

GGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTG

GCCTTTTGCTGGCCTTTTGCTCACATGT pGW07 1-pAA V_EFSLTR-Cd80-T2 A-light-P2 A-cxcl 10-E2 A-41 bb-F2 A-I12-sPA (SEQ ID NO: 24)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgTtgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctgggctggccactc agggctggtttctcctgagactgca tcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggagaa gctgatacaagatcaacgatctca ccaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtgg acctctgttatgggagacacgact tggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagcc cggttactactatgtgtactccaaag tgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccatg gactatacaagcgcacatcccg ctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaacag ctcccgagtctggtgggacagca gcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcctg gaaaccgcctggtcagaccacgt gacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGCAACAAAC TTCTCTCTG

CTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgaacccaagtgctgcc gtc attttctgcctcatcctgctgggtctgagtgggactcaagggatccctctcgcaaggacg gtccgctgcaactgcatccatatcgat gacgggccagtgagaatgagggccatagggaagcttgaaatcatccctgcgagcctatcc tgcccacgtgttgagatcattgcc acgatgaaaaagaatgatgagcagagatgtctgaatccggaatctaagaccatcaagaat ttaatgaaagcgtttagccaaaaaa ggtctaaaagggctcctgctagcagcggtacccagtgcaccaactacgccctgctgaagc tggccggcgatgtggagagcaa ccccgggcccatggaccagcacacacttgatgtggaggataccgcggatgccagacatcc agcaggtacttcgtgcccctcgg atgcggcgctcctcagagataccgggctcctcgcggacgctgcgctcctctcagatactg tgcgccccacaaatgccgcgctcc ccacggatgctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctg cactgaacttctgttcccgccaccc aaagctctatggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctat cttcacccgcaccgagcctcggccagcg ctcacaatcaccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacc cctgtttcccacattggctgcccca acactacacaacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgt tgtgcaatacaactctgaactggcac agccaagatggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaa aaggagttggtggtagacagtcc cgggctctactacgtatttttggaactgaagctcagtccaacattcacaaacacaggcca caaggtgcagggctgggtctctcttg ttttgcaagcaaagcctcaggtagatgactttgacaacttggccctgacagtggaactgt tcccttgctccatggagaacaagttag tggaccgttcctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtc tgagggcttatctgcatggagccca ggatgcatacagagactgggagctgtcttatcccaacaccaccagctttggactctttct tgtgaaacccgacaacccatgggaa ggcatatgcggtaccgtgaagcagaccctgaacttcgatctgctgaagctggccggcgat gtggagagcaaccccgggcccat gtacagcatgcagctcgcatcctgtgtcacattgacacttgtgctccttgtcaacagcgc acccacttcaagctccacttcaagctct acagcggaagcacagcagcagcagcagcagcagcagcagcagcagcagcacctggagcag ctgttgatggacctacagga gctcctgagcaggatggagaattacaggaacctgaaactccccaggatgctcaccttcaa attttacttgcccaagcaggccaca gaattgaaagatcttcagtgcctagaagatgaacttggacctctgcggcatgttctggat ttgactcaaagcaaaagctttcaattgg aagatgctgagaatttcatcagcaatatcagagtaactgttgtaaaactaaagggctctg acaacacatttgagtgccaattcgatg atgagtcagcaactgtggtggactttctgaggagatggatagccttctgtcaaagcatca tctcaacaagccctcaataaGAAT

TCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGG

CCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTC

GCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGG

GCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGC

GGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGC

AACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGT

TACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCG

CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAA

ATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCC

AAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGA

CGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGT

TCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAA

GGGATTTTGCCGATTTCGGTCT ATT GGTT AAA AAAT GAGCTGATTT A AC AAAA

ATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTC

TCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCA

ACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAG

ACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

CACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT

TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAA

ATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC

CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA

GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT

TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG

TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT

TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAG

CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC

AGT C AC AGAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGA ATT AT GC AGT

GCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT

CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA

ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG

AGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT

AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG

AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG

GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTG

CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGAC

GGGGAGTC AGGC A ACT AT GGAT GA ACGA A AT AGAC AGATCGC T GAG AT AGG

TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATAC

TTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATC

CTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA

GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCT

GCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTT

TGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAG

CAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACC

ACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA

CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW072-pAAV EFSLTR-IFNg-sPA (SEQ ID NO: 25)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgctgaGAATTCAATAAAAGATCTTTATTTT CATTA GATCTGTGT GTT GGTTTTTTGT GT GCGGCCGC AGGAACCCCT AGT GAT GGAGT TGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAG GTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGC GCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGC GGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGG CGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTT GCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACG TTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCG ATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTT CACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAG TCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTC TATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTG GTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATA TTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGC ATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGG GCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAG CTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAG GGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCT TAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTT ATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGAT AAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCG TGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCA GAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGG GTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCC GAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGT ATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATT CTC AGA AT GAC TT GGTT GAGT ACTC AC C AGT C AC AGA A A AGC AT C TT ACGGA T GGC AT G AC AGT A AG AG A AT TAT GC AGT GC T GC CAT A AC CAT G AGT GAT A AC ACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCG _{CTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCG} GAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAG CAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCT TCCCGGC AAC AATT AAT AGACTGGAT GGAGGCGGAT AAAGTT GC AGGACC AC TTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCC GGT GAGCGTGGAAGCCGCGGT ATC ATT GC AGC ACTGGGGCC AGAT GGT A AGC CCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGA ACGAAAT AGAC AGATCGCTGAGAT AGGT GCCTC ACTGATT AAGC ATTGGT AA CTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTT TAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAAT CCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCA AAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAA AAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAAC TCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTC TTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCT ACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAA GTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAG CGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGA CCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCT TCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAAC

AGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGT

CCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCA

GGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC

TGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW073-pAA V_EFSLTR-Cd80-T2 A-light-P2 A-cxcl 10-E2 A-41 bb-F2 A-Gitrl (SEQ ID NO: 26)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgTtgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctgggctggccactc agggctggtttctcctgagactgca tcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggagaa gctgatacaagatcaacgatctca ccaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtgg acctctgttatgggagacacgact tggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagcc cggttactactatgtgtactccaaag tgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccatg gactatacaagcgcacatcccg ctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaacag ctcccgagtctggtgggacagca gcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcctg gaaaccgcctggtcagaccacgt gacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGCAACAAAC TTCTCTCTG

CTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgaacccaagtgctgcc gtc attttctgcctcatcctgctgggtctgagtgggactcaagggatccctctcgcaaggacg gtccgctgcaactgcatccatatcgat gacgggccagtgagaatgagggccatagggaagcttgaaatcatccctgcgagcctatcc tgcccacgtgttgagatcattgcc acgatgaaaaagaatgatgagcagagatgtctgaatccggaatctaagaccatcaagaat ttaatgaaagcgtttagccaaaaaa ggtctaaaagggctcctgctagcagcggtacccagtgcaccaactacgccctgctgaagc tggccggcgatgtggagagcaa ccccgggcccatggaccagcacacacttgatgtggaggataccgcggatgccagacatcc agcaggtacttcgtgcccctcgg atgcggcgctcctcagagataccgggctcctcgcggacgctgcgctcctctcagatactg tgcgccccacaaatgccgcgctcc ccacggatgctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctg cactgaacttctgttcccgccaccc aaagctctatggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctat cttcacccgcaccgagcctcggccagcg ctcacaatcaccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacc cctgtttcccacattggctgcccca acactacacaacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgt tgtgcaatacaactctgaactggcac agccaagatggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaa aaggagttggtggtagacagtcc cgggctctactacgtatttttggaactgaagctcagtccaacattcacaaacacaggcca caaggtgcagggctgggtctctcttg ttttgcaagcaaagcctcaggtagatgactttgacaacttggccctgacagtggaactgt tcccttgctccatggagaacaagttag tggaccgttcctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtc tgagggcttatctgcatggagccca ggatgcatacagagactgggagctgtcttatcccaacaccaccagctttggactctttct tgtgaaacccgacaacccatgggaa ggcatatgcggtaccgtgaagcagaccctgaacttcgatctgctgaagctggccggcgat gtggagagcaaccccgggcccat ggaggaaatgcctttgagagagtcaagtcctcaaagggcagagaggtgcaagaagtcatg gctcttgtgcatagtggctctgtta ctgatgctgctctgttctttgggtacactgatctatacttcactcaagccaactgccatc gagtcctgcatggttaagtttgaactatca tcctcaaaatggcacatgacatctcccaaacctcactgtgtgaatacgacatctgatggg aagctgaagatactgcagagtggca catatttaatctacggccaagtgattcctgtggataagaaatacataaaagacaatgccc ccttcgtagtacagatatataaaaaga atgatgtcctacaaactctaatgaatgattttcaaatcttgcctataggaggggtttatg aactgcatgctggagataacatatatctg aagttcaactctaaagaccatattcagaaaaataacacatactgggggatcatcttaatg cctgatctaccattcatctcttagGAA

TTCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCG

GCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCT

CGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCG

GGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATG

CGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAG

CAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGG

TTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTC

GCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTA

AATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCC

CAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAG

ACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTG

TTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATA

AGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAA

AATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACT

CTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCC

AACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACA

GACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCA

TCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGG

TTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGA

AATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTAT

CCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGA

AGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCAT

TTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCT

GAAGATC AGTT GGGTGC ACGAGTGGGTT AC ATCGAACTGGATCTC AAC AGCG

GTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACT

TTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGA

GCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCAC

C AGT C AC AGA A A AGC ATCTT ACGG AT GGC AT G AC AGT A AG AG A ATT AT GC AG

TGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGA

TCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGT

AACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGAC

GAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTAT

TAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATG

GAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCT

GGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATT GCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGA

CGGGGAGT C AGGC A ACT AT GGAT GA AC GA A AT AGAC AG AT C GCTGAGAT AG

GTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATA

CTTT AGATTGATTT AAAACTT C ATTTTT AATTT AAAAGGATCT AGGT GAAGAT

CCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTG

AGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTC

TGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGT

TTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCA

GCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCAC

CACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTT

ACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAA

GACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTG

CACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAG

CGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGG

TATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCA

GGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACT

TGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAAC

GCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCAC

ATGT pGW078-pAAV EFSLTR-Agel&EcoRI-sPA (SEQ ID NO: 27)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c tgaGAATTCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGT

GTGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCG

CTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTT

TGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGC

CTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGT

CAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGT

GGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTC

CTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAG

CTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTC

GACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTG

ATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGAC

TCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATT

TATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAA

CAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTG

CACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACC

CGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCT TACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACC

GTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTA

TAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCG

GGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATA

TGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAA

AGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCG

GCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGA

T GCTGAAGATC AGTT GGGT GC ACGAGTGGGTT AC ATCGAACTGGATCTC AAC

AGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAG

CACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGC

AAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTAC

T C ACC AGT C AC AGAAAAGC ATCTT ACGGAT GGC AT GAC AGT AAGAGAATT AT

GCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACA

ACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATC

ATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAA

CGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAA

CTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTG

GATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCT

GGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTA

TCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTAC

ACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAG

ATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATA

TATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGA

AGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCC

ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTT

TTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGT

GGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCT

TCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGC

CACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCT

GTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACT

CAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTC

GTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTA

CAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGAC

AGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTT

CCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTG

ACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

AACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCT

CACATGT pGW090-pAAV EFSLTR-Cd80-T2A-light-P2A-IL2-E2A-41bb-F2A-Ifng (SEQ ID NO: 28)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgTtgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctgggctggccactc agggctggtttctcctgagactgca tcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggagaa gctgatacaagatcaacgatctca ccaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtgg acctctgttatgggagacacgact tggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagcc cggttactactatgtgtactccaaag tgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccatg gactatacaagcgcacatcccg ctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaacag ctcccgagtctggtgggacagca gcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcctg gaaaccgcctggtcagaccacgt gacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGCAACAAAC TTCTCTCTG

CTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgtacagcatgcagctc gca tcctgtgtcacattgacacttgtgctccttgtcaacagcgcacccacttcaagctccact tcaagctctacagcggaagcacagca gcagcagcagcagcagcagcagcagcagcagcacctggagcagctgttgatggacctaca ggagctcctgagcaggatgga gaattacaggaacctgaaactccccaggatgctcaccttcaaattttacttgcccaagca ggccacagaattgaaagatcttcagt gcctagaagatgaacttggacctctgcggcatgttctggatttgactcaaagcaaaagct ttcaattggaagatgctgagaatttcat cagcaatatcagagtaactgttgtaaaactaaagggctctgacaacacatttgagtgcca attcgatgatgagtcagcaactgtggt ggactttctgaggagatggatagccttctgtcaaagcatcatctcaacaagccctcaagc tagcagcggtacccagtgcaccaac tacgccctgctgaagctggccggcgatgtggagagcaaccccgggcccatggaccagcac acacttgatgtggaggataccg cggatgccagacatccagcaggtacttcgtgcccctcggatgcggcgctcctcagagata ccgggctcctcgcggacgctgcg ctcctctcagatactgtgcgccccacaaatgccgcgctccccacggatgctgcctaccct gcggttaatgttcgggatcgcgagg ccgcgtggccgcctgcactgaacttctgttcccgccacccaaagctctatggcctagtcg ctttggttttgctgcttctgatcgccgc ctgtgttcctatcttcacccgcaccgagcctcggccagcgctcacaatcaccacctcgcc caacctgggtacccgagagaataat gcagaccaggtcacccctgtttcccacattggctgccccaacactacacaacagggctct cctgtgttcgccaagctactggcta aaaaccaagcatcgttgtgcaatacaactctgaactggcacagccaagatggagctggga gctcatacctatctcaaggtctgag gtacgaagaagacaaaaaggagttggtggtagacagtcccgggctctactacgtattttt ggaactgaagctcagtccaacattca caaacacaggccacaaggtgcagggctgggtctctcttgttttgcaagcaaagcctcagg tagatgactttgacaacttggccct gacagtggaactgttcccttgctccatggagaacaagttagtggaccgttcctggagtca actgttgctcctgaaggctggccacc gcctcagtgtgggtctgagggcttatctgcatggagcccaggatgcatacagagactggg agctgtcttatcccaacaccacca gctttggactctttcttgtgaaacccgacaacccatgggaaggcatatgcggtaccgtga agcagaccctgaacttcgatctgctg aagctggccggcgatgtggagagcaaccccgggcccatgaacgctacacactgcatcttg gctttgcagctcttcctcatggctg tttctggctgttactgccacggcacagtcattgaaagcctagaaagtctgaataactatt ttaactcaagtggcatagatgtggaaga aaagagtctcttcttggatatctggaggaactggcaaaaggatggtgacatgaaaatcct gcagagccagattatctctttctacct cagactctttgaagtcttgaaagacaatcaggccatcagcaacaacataagcgtcattga atcacacctgattactaccttcttcag caacagcaaggcgaaaaaggatgcattcatgagtattgccaagtttgaggtcaacaaccc acaggtccagcgccaagcattcaa tgagctcatccgagtggtccaccagctgttgccggaatccagcctcaggaagcggaaaag gagtcgctgctgaGAATTC AATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCC

GCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGC

TCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGC

GGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGG

TATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAA

CCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTA

CGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCT

TTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAAT

CGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAA

AAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACG

GTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTC

CAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGG

GATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAAT

TTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTC

AGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAA

CACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGA

CAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATC

ACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTT

AATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAA

TGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC

GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAG

AGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTT

TGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGA

AGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGT

AAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTT

TAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGC

AACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCA

GT C AC AGA A A AGC AT C TT ACGGAT GGC AT GAC AGT A AG AG A ATT AT GC AGT G

CTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATC

GGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAA

CTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGA

GCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTA

ACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGA

GGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGT

TTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGC

AGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACG

GGGAGTC AGGC AACT AT GGAT GAACGAA AT AGAC AGATCGCTGAGAT AGGT

GCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACT

TTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCC

TTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAG

CGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTG

CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTT

GTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC

AGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCA

CTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTAC

CAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW091-pAAV EFSLTR-Cd80-T2A-light-P2A-GITRL-E2A-41bb-F2A-Ifng (SEQ ID NO: 29)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgTtgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctgggctggccactc agggctggtttctcctgagactgca tcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggagaa gctgatacaagatcaacgatctca ccaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtgg acctctgttatgggagacacgact tggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagcc cggttactactatgtgtactccaaag tgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccatg gactatacaagcgcacatcccg ctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaacag ctcccgagtctggtgggacagca gcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcctg gaaaccgcctggtcagaccacgt gacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGCAACAAAC TTCTCTCTG

CTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggaggaaatgcctttg aga gagtcaagtcctcaaagggcagagaggtgcaagaagtcatggctcttgtgcatagtggct ctgttactgatgctgctctgttctttg ggtacactgatctatacttcactcaagccaactgccatcgagtcctgcatggttaagttt gaactatcatcctcaaaatggcacatga catctcccaaacctcactgtgtgaatacgacatctgatgggaagctgaagatactgcaga gtggcacatatttaatctacggccaa gtgattcctgtggataagaaatacataaaagacaatgcccccttcgtagtacagatatat aaaaagaatgatgtcctacaaactcta atgaatgattttcaaatcttgcctataggaggggtttatgaactgcatgctggagataac atatatctgaagttcaactctaaagacca tattcagaaaaataacacatactgggggatcatcttaatgcctgatctaccattcatctc tgctagcagcggtacccagtgcaccaa ctacgccctgctgaagctggccggcgatgtggagagcaaccccgggcccatggaccagca cacacttgatgtggaggatacc gcggatgccagacatccagcaggtacttcgtgcccctcggatgcggcgctcctcagagat accgggctcctcgcggacgctgc gctcctctcagatactgtgcgccccacaaatgccgcgctccccacggatgctgcctaccc tgcggttaatgttcgggatcgcgag gccgcgtggccgcctgcactgaacttctgttcccgccacccaaagctctatggcctagtc gctttggttttgctgcttctgatcgcc gcctgtgttcctatcttcacccgcaccgagcctcggccagcgctcacaatcaccacctcg cccaacctgggtacccgagagaat aatgcagaccaggtcacccctgtttcccacattggctgccccaacactacacaacagggc tctcctgtgttcgccaagctactgg ctaaaaaccaagcatcgttgtgcaatacaactctgaactggcacagccaagatggagctg ggagctcatacctatctcaaggtct gaggtacgaagaagacaaaaaggagttggtggtagacagtcccgggctctactacgtatt tttggaactgaagctcagtccaaca ttcacaaacacaggccacaaggtgcagggctgggtctctcttgttttgcaagcaaagcct caggtagatgactttgacaacttggc cctgacagtggaactgttcccttgctccatggagaacaagttagtggaccgttcctggag tcaactgttgctcctgaaggctggcc accgcctcagtgtgggtctgagggcttatctgcatggagcccaggatgcatacagagact gggagctgtcttatcccaacaccac cagctttggactctttcttgtgaaacccgacaacccatgggaaggcatatgcggtaccgt gaagcagaccctgaacttcgatctgc tgaagctggccggcgatgtggagagcaaccccgggcccatgaacgctacacactgcatct tggctttgcagctcttcctcatggc tgtttctggctgttactgccacggcacagtcattgaaagcctagaaagtctgaataacta ttttaactcaagtggcatagatgtggaa gaaaagagtctcttcttggatatctggaggaactggcaaaaggatggtgacatgaaaatc ctgcagagccagattatctctttctac ctcagactctttgaagtcttgaaagacaatcaggccatcagcaacaacataagcgtcatt gaatcacacctgattactaccttcttca gcaacagcaaggcgaaaaaggatgcattcatgagtattgccaagtttgaggtcaacaacc cacaggtccagcgccaagcattca atgagctcatccgagtggtccaccagctgttgccggaatccagcctcaggaagcggaaaa ggagtcgctgctgaGAATTC

AATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCC

GCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGC

TCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGC

GGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGG

TATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAA

CCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTA

CGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCT

TTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAAT

CGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAA

AAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACG

GTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTC

CAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGG

GATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAAT

TTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTC

AGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAA

CACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGA

CAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATC

ACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTT

AATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAA

TGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC

GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAG

AGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTT

TGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGA

AGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGT

AAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTT

TAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGC

AACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCA

GT C AC AGA A A AGC AT C TT ACGGAT GGC AT GAC AGT A AG AG A ATT AT GC AGT G

CTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATC

GGAGGACCGA AGGAGCT AACCGCTTTTTTGC AC AAC AT GGGGGAT CAT GT AA

CTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGA

GCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTA

ACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGA

GGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGT TTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGC

AGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACG

GGGAGTC AGGC AACT AT GGAT GAACGAA AT AGAC AGATCGCTGAGAT AGGT

GCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACT

TTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCC

TTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAG

CGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTG

CGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTT

GTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC

AGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCA

CTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTAC

CAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW096-pAAV EFSLTR-Ifng-P2A-Cd80-T2A-light-P2A-IL23-E2A-41BBL-sPA (SEQ ID NO: 30)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGC AGTGGAGAGGGC AGAGGAAGTCTGCT AAC AT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctgc cagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcca acccagcagcacatcttacagg agccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcct ggccttcttgaggggcttgacgtatc atgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagc tgagcggcgtgggctgcccccagg ggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccga aggagttagaactgctggtcagtc ggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgg gcggcgtggtacatctggaggc tggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcac caggtcctatttcggagctttcat ggtcactagtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATG

TCGAAGAGAATCCTGGACCGATGTGTCCTCAGAAGCTAACCATCTCCTGGTTT

GCCATCGTTTTGCTGGTGTCTCCACTCATGGCCATGTGGGAGCTGGAGAAAGA

CGTTT AT GTT GT AGAGGTGGACTGGACTCCCGAT GCCCCTGGAGAAAC AGT G

AACCTCACCTGTGACACGCCTGAAGAAGATGACATCACCTGGACCTCAGACC

AGAGACATGGAGTCATAGGCTCTGGAAAGACCCTGACCATCACTGTCAAAGA

GTTTCTAGATGCTGGCCAGTACACCTGCCACAAAGGAGGCGAGACTCTGAGC

CACTCACATCTGCTGCTCCACAAGAAGGAAAATGGAATTTGGTCCACTGAAA

TTTTAAAAAATTTCAAAAACAAGACTTTCCTGAAGTGTGAAGCACCAAATTAC

TCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGT

TCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGG

AATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAG

AAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAGA

CCCTGCCC ATT GAACTGGCGTTGGAAGC ACGGC AGC AGAAT AAAT AT GAGAA

CTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAG

AACTTGC AGAT GAAGCCTTTGAAGAACTC AC AGGT GGAGGT C AGCTGGGAGT

ACCCTGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTC

GAATCCAGCGCAAGAAAGAAAAGATGAAGGAGACAGAGGAGGGGTGTAACC

AGAAAGGTGCGTTCCTCGT AGAGAAGAC ATCT ACCGAAGTCC AAT GC AA AGG

CGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCA

AGTGGGCATGTGTTCCCTGCAGGGTCCGATCCgctagcagcggtacccagtgcacca actacgc cctgctgaagctggccggcgatgtggagagcaaccccgggcccatggaccagcacacact tgatgtggaggataccgcggat gccagacatccagcaggtacttcgtgcccctcggatgcggcgctcctcagagataccggg ctcctcgcggacgctgcgctcct ctcagatactgtgcgccccacaaatgccgcgctccccacggatgctgcctaccctgcggt taatgttcgggatcgcgaggccgc gtggccgcctgcactgaacttctgttcccgccacccaaagctctatggcctagtcgcttt ggttttgctgcttctgatcgccgcctgt gttcctatcttcacccgcaccgagcctcggccagcgctcacaatcaccacctcgcccaac ctgggtacccgagagaataatgca gaccaggtcacccctgtttcccacattggctgccccaacactacacaacagggctctcct gtgttcgccaagctactggctaaaaa ccaagcatcgttgtgcaatacaactctgaactggcacagccaagatggagctgggagctc atacctatctcaaggtctgaggtac gaagaagacaaaaaggagttggtggtagacagtcccgggctctactacgtatttttggaa ctgaagctcagtccaacattcacaa acacaggccacaaggtgcagggctgggtctctcttgttttgcaagcaaagcctcaggtag atgactttgacaacttggccctgac agtggaactgttcccttgctccatggagaacaagttagtggaccgttcctggagtcaact gttgctcctgaaggctggccaccgcc tcagtgtgggtctgagggcttatctgcatggagcccaggatgcatacagagactgggagc tgtcttatcccaacaccaccagcttt ggactctttcttgtgaaacccgacaacccatgggaatgaGAATTCAATAAAAGATCTTTA TTTTCATT

AGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCCCTAGTGATGGAG

TTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAA

GGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCG

CGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTG CGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCG

GCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACT

TGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCAC

GTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCC

GATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGT

TCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGA

GTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACT

CTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATT

GGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAAT

ATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCG

CATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACG

GGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGA

GCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAA

GGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTT

CTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGT

TTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTG

AT A A AT GCTT C A AT A AT ATT GA A A A AGG A AGAGT AT GAGT ATTC A AC ATTT C

CGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACC

C AG A A AC GC T GGT G A A AGT A A A AG AT GC T G A AG AT C AGT T GGGT GC AC GAGT

GGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCC

CCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCG

GTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACT

ATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACG

GAT GGC AT GAC AGT A AG AG A ATT AT GC AGT GCTGCC AT A AC CAT GAGT GAT A

ACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAAC

CGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAAC

CGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGT

AGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA

GCTTCCCGGC AAC AATT AAT AGACTGGAT GGAGGCGGAT AA AGTT GC AGGAC

CACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGA

GCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTA

AGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGA

TGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGG

TAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCA

TTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA

AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAG

ATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAA

ACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTAC

CAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACT

GTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC

GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCG

ATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGC

GCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGA

ACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCA

CGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCG

GAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTA

TAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTC

GTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG

TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW097-pAAV EFSLTR-IL23-sPA (SEQ ID NO: 31)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac

ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGCTGGTGTCT

CCACTCATGGCCATGTGGGAGCTGGAGAAAGACGTTTATGTTGTAGAGGTGG

ACTGGACTCCCGATGCCCCTGGAGAAACAGTGAACCTCACCTGTGACACGCC

TGAAGAAGATGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGC

TCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTTCTAGATGCTGGCCAGTA

CACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCAC

AAGAAGGAAAAT GGAATTT GGTCC ACTGAAATTTT AAAAAATTT C AAAA AC A

AGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCA

T GGCTGGTGC A AAGAAAC ATGGACTTGAAGTTC AAC AT C AAGAGC AGT AGC A

GTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAG

AAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGG

AGGATGTCACCTGCCCAACTGCCGAGGAGACCCTGCCCATTGAACTGGCGTT

GGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATC

AGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGA

AGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCTGACTCCTGGAGCACTCC

CCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAAA

AG AT G A AGG AG AC AG AGG AGGGGT GT AAC C AG A A AGGT GC GT TC C T C GT AG

AGAAGAC ATCT ACCGAAGTCC AATGC AAAGGCGGGAATGTCTGCGT GCA AGC

TCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCA

GGGTCCGATCCTAGGAATTCAATAAAAGATCTTTATTTTCATTAGATCTGTGT

GTTGGTTTTTTGTGTGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTC

CCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCG

ACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGC

CTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTC

ACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAA

GCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGC

CTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGG

CTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG

CTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGT

GGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTT

CTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGG

GCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAA

ATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTT

ACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAG

CCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTG

CTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTG TCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTG

ATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTC

AGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCT

AAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTT

C AAT AAT ATT GAAAAAGGA AGAGT ATGAGT ATT C A AC ATTTCCGT GTCGCCCT

TATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCT

GGT GA A AGT A A A AG AT GCTGA AGAT C AGTT GGGT GC ACGAGT GGGTT AC AT C

GAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAAC

GTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCC

CGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGA

ATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT

GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCG

GCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTT

GCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTG

AATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGG

CAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGG

C A AC A ATT A AT AGACTGGAT GGAGGC GGAT A A AGTT GC AGG ACC ACTT C T GC

GCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAG

CGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCC

GTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAA

TAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCA

GACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTT

AAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTA

ACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGA

TCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA

CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTT

TCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAG

TGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATAC

CTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTG

TCTT ACCGGGTT GGACTC AAGACGAT AGTT ACCGGAT AAGGCGC AGCGGTCG

GGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACA

CCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGA

AGGGAGA A AGGC GGAC AGGT ATCCGGT A AGC GGC AGGGTCGGA AC AGGAGA

GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTC

GGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGG

GCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCC

TTTTGCTGGCCTTTTGCTCACATGT pGW098-pAAV EFSLTR-Ifng-P2A-Cd80-T2A-light-P2A-IL23-sPA (SEQ ID NO: 32)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGCAGTGGAGAGGGCAGAGGAAGTCTGCTAACAT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctgc cagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcca acccagcagcacatcttacagg agccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcct ggccttcttgaggggcttgacgtatc atgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagc tgagcggcgtgggctgcccccagg ggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccga aggagttagaactgctggtcagtc ggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgg gcggcgtggtacatctggaggc tggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcac caggtcctatttcggagctttcat ggtcactagtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATG

TCGAAGAGAATCCTGGACCGATGTGTCCTCAGAAGCTAACCATCTCCTGGTTT

GCCATCGTTTTGCTGGTGTCTCCACTCATGGCCATGTGGGAGCTGGAGAAAGA

CGTTT AT GTT GT AGAGGTGGACTGGACTCCCGAT GCCCCTGGAGAAAC AGT G

AACCTCACCTGTGACACGCCTGAAGAAGATGACATCACCTGGACCTCAGACC

AGAGACATGGAGTCATAGGCTCTGGAAAGACCCTGACCATCACTGTCAAAGA

GTTTCTAGATGCTGGCCAGTACACCTGCCACAAAGGAGGCGAGACTCTGAGC

CACTCACATCTGCTGCTCCACAAGAAGGAAAATGGAATTTGGTCCACTGAAA

TTTTAAAAAATTTCAAAAACAAGACTTTCCTGAAGTGTGAAGCACCAAATTAC

TCCGGACGGTTCACGTGCTCATGGCTGGTGCAAAGAAACATGGACTTGAAGT

TCAACATCAAGAGCAGTAGCAGTTCCCCTGACTCTCGGGCAGTGACATGTGG

AATGGCGTCTCTGTCTGCAGAGAAGGTCACACTGGACCAAAGGGACTATGAG

AAGTATTCAGTGTCCTGCCAGGAGGATGTCACCTGCCCAACTGCCGAGGAGA

CCCTGCCC ATT GAACTGGCGTTGGAAGC ACGGC AGC AGAAT AAAT AT GAGAA

CTACAGCACCAGCTTCTTCATCAGGGACATCATCAAACCAGACCCGCCCAAG

AACTTGC AGAT GAAGCCTTTGAAGAACTC AC AGGT GGAGGT C AGCTGGGAGT

ACCCTGACTCCTGGAGCACTCCCCATTCCTACTTCTCCCTCAAGTTCTTTGTTC

GAATCCAGCGCAAGAAAGAAAAGATGAAGGAGACAGAGGAGGGGTGTAACC

AGAAAGGTGCGTTCCTCGT AGAGAAGAC ATCT ACCGAAGTCC AAT GC AA AGG

CGGGAATGTCTGCGTGCAAGCTCAGGATCGCTATTACAATTCCTCATGCAGCA AGTGGGCATGTGTTCCCTGCAGGGTCCGATCCtgaGAATTCAATAAAAGATCTT

TATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCCCTA

GTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGG

GCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGC

GAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTAC

GCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCC

CTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACC

GCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTT

CTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTT

AGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGG

GTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTG

ACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAAC

ACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTC

GGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTA

ACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCT

GATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGC

CCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTC

TCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGA

GACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATA

ATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCC

TATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATA

ACC CTGAT A A AT GC TT C A AT A AT ATTGA A A A AGGA AGAGT AT GAGT ATT C A A

CATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTG

CTC ACCC AGAAACGCTGGTGAAAGT AAAAGAT GCTGAAGATC AGTT GGGT GC

ACGAGTGGGTT AC ATCGAACTGGATCTC AAC AGCGGT AAGATCCTTGAGAGT

TTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATG

TGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGC

AT AC ACT ATTCTC AGAAT GACTT GGTTGAGT ACTC ACC AGT C AC AGAAAAGC

ATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCAT

GAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAG

GAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCG

TTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACG

AT GCCTGT AGC AAT GGC AAC AACGTTGCGC AAACT ATT AACTGGCGA ACT AC

TTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGT

TGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATA

AATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCC

AGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCA

AC T AT GG AT G A AC G A A AT AG AC AG AT C GC T GAG AT AGGT GC C TC AC T GATT A

AGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTA

AAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCT

CATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCG

TAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGC

TGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATC

AAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGAT

ACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACT

CTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCT

GCC AGT GGC GAT AAGTCGT GTCTT ACCGGGTT GGACTC AAGACGAT AGTT AC

CGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAG

CTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGA

GAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGC GGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCC

TGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTT

TTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGG

CCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW099-pAAV EFSLTR-IFNg-P2A-Cd80-T2A-light-P2A-cxcll0 (SEQ ID NO: 33)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGCAGTGGAGAGGGCAGAGGAAGTCTGCTAACAT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctgc cagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcca acccagcagcacatcttacagg agccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcct ggccttcttgaggggcttgacgtatc atgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagc tgagcggcgtgggctgcccccagg ggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccga aggagttagaactgctggtcagtc ggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgg gcggcgtggtacatctggaggc tggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcac caggtcctatttcggagctttcat ggtcactagtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATG

TCGAAGAGAATCCTGGACCGatgaacccaagtgctgccgtcattttctgcctcatcc tgctgggtctgagtggg actcaagggatccctctcgcaaggacggtccgctgcaactgcatccatatcgatgacggg ccagtgagaatgagggccatagg gaagcttgaaatcatccctgcgagcctatcctgcccacgtgttgagatcattgccacgat gaaaaagaatgatgagcagagatgt ctgaatccggaatctaagaccatcaagaatttaatgaaagcgtttagccaaaaaaggtct aaaagggctccttgaGAATTCA

ATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCG

CAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCT

CACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCG

GCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGT

ATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAAC

CATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTAC

GCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTT

TCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATC

GGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAA

AAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGT

TTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCA

AACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGA

TTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTT

AACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAG

TACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACA

CCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACA

AGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCAC

CGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAA

TGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATG

TGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGC

T CAT GAGAC AAT AACCCTGAT AAAT GCTTC AAT AAT ATTGAAAAAGGAAGAG

TATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTG

CCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAG

AT C AGTTGGGT GC ACGAGT GGGTT AC ATCGAACTGGATCTC AAC AGCGGT AA

GATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTA

AAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCA

ACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAG

T C AC AG A A A AGC AT C TT ACGGAT GGC AT GAC AGT A AG AG A ATT AT GC AGT GC

TGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCG

GAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAAC

TCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAG

CGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAA

CTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAG

GCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTT

TATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCA

GCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGG

GGAGT C AGGC A AC T AT GGAT G A ACGA A AT AGAC AGATCGC T GAG AT AGGT GC

CTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTT

AGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTT

TTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGC

GTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGC

GCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTG

TTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCA

GAGCGCAGAT ACC AAAT ACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCAC

TTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACC

AGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGA

CGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCA

CACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCG

TGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTA TCCGGT AAGCGGC AGGGTCGGAAC AGGAGAGCGC ACGAGGGAGCTTCC AGG

GGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTG

AGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGC

CAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACAT

GT pGWIOO-pAAV EFSLTR-Ifng-P2A-Cd80-T2A-light-P2A-Cxcll0-E2A-41BBL-sPA (SEQ ID NO: 34)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGC AGTGGAGAGGGC AGAGGAAGTCTGCT AAC AT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaAgtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctg ccagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcc aacccagcagcacatcttacag gagccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcc tggccttcttgaggggcttgacgtat catgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcag ctgagcggcgtgggctgcccccag gggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccg aaggagttagaactgctggtcagt cggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctg ggcggcgtggtacatctggagg ctggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggca ccaggtcctatttcggagctttca tggtcactagtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATG

TCGAAGAGAATCCTGGACCGatgaacccaagtgctgccgtcattttctgcctcatcc tgctgggtctgagtggg actcaagggatccctctcgcaaggacggtccgctgcaactgcatccatatcgatgacggg ccagtgagaatgagggccatagg gaagcttgaaatcatccctgcgagcctatcctgcccacgtgttgagatcattgccacgat gaaaaagaatgatgagcagagatgt ctgaatccggaatctaagaccatcaagaatttaatgaaagcgtttagccaaaaaaggtct aaaagggctcctgctagcagcggta cccagtgcaccaactacgccctgctgaagctggccggcgatgtggagagcaaccccgggc ccatggaccagcacacacttga tgtggaggataccgcggatgccagacatccagcaggtacttcgtgcccctcggatgcggc gctcctcagagataccgggctcct cgcggacgctgcgctcctctcagatactgtgcgccccacaaatgccgcgctccccacgga tgctgcctaccctgcggttaatgtt cgggatcgcgaggccgcgtggccgcctgcactgaacttctgttcccgccacccaaagctc tatggcctagtcgctttggttttgct gcttctgatcgccgcctgtgttcctatcttcacccgcaccgagcctcggccagcgctcac aatcaccacctcgcccaacctgggt acccgagagaataatgcagaccaggtcacccctgtttcccacattggctgccccaacact acacaacagggctctcctgtgttcg ccaagctactggctaaaaaccaagcatcgttgtgcaatacaactctgaactggcacagcc aagatggagctgggagctcatacc tatctcaaggtctgaggtacgaagaagacaaaaaggagttggtggtagacagtcccgggc tctactacgtatttttggaactgaag ctcagtccaacattcacaaacacaggccacaaggtgcagggctgggtctctcttgttttg caagcaaagcctcaggtagatgactt tgacaacttggccctgacagtggaactgttcccttgctccatggagaacaagttagtgga ccgttcctggagtcaactgttgctcct gaaggctggccaccgcctcagtgtgggtctgagggcttatctgcatggagcccaggatgc atacagagactgggagctgtctta tcccaacaccaccagctttggactctttcttgtgaaacccgacaacccatgggaatgaGA ATTCAATAAAAGATC

TTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCCC

TAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCC

GGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGA

GCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTT

ACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGC

GCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTG

ACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCC

TTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCC

TTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATT

TGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCT

TTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAAC

AACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGAT

TTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATT

TTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGC

TCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACG

CGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACC

GTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGC

GAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAA

TAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACC

CCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAA

T A ACCC T GAT A A AT GCTT C A AT A AT ATT GA A A A AGGA AGAGT AT GAGT ATTC

AACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTT

T GCTC ACCC AGAAACGCTGGTGAA AGT A AAAGAT GCTGAAGAT C AGTTGGGT

GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGA

GTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTA

TGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCC

GCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA

GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACC

ATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA

AGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGAT

CGTTGGGAACCGGAGCTGAAT GAAGCC AT ACC AAACGACGAGCGT GAC ACC A

CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACT

ACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAA

GTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGA

TAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGG CCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG

CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGAT

TAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATT

TAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAAT

CTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCC

CGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCT

GCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGA

TCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAG

ATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA

CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTG

CTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTA

CCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCA

GCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATG

AGAAAGCGCC ACGCTTCCCGAAGGGAGAAAGGCGGAC AGGT ATCCGGT AAG

CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGC

CTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATT

TTTGT GAT GCTCGT C AGGGGGGCGGAGCCT AT GGAAAAACGCC AGC AACGCG

GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGWIOl-pAAV EFSLTR-Ifng-P2A-Cd80-T2A-light-E2A-41BBL-sPA (SEQ ID NO: 35)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGC AGTGGAGAGGGC AGAGGAAGTCTGCT AAC AT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctgc cagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcca acccagcagcacatcttacagg agccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcct ggccttcttgaggggcttgacgtatc atgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagc tgagcggcgtgggctgcccccagg ggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccga aggagttagaactgctggtcagtc ggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgg gcggcgtggtacatctggaggc tggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcac caggtcctatttcggagctttcat ggtcactagcagcggtacccagtgcaccaactacgccctgctgaagctggccggcgatgt ggagagcaaccccgggcccatg gaccagcacacacttgatgtggaggataccgcggatgccagacatccagcaggtacttcg tgcccctcggatgcggcgctcct cagagataccgggctcctcgcggacgctgcgctcctctcagatactgtgcgccccacaaa tgccgcgctccccacggatgctg cctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctgcactgaacttct gttcccgccacccaaagctctatgg cctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctatcttcacccgcac cgagcctcggccagcgctcacaatcacc acctcgcccaacctgggtacccgagagaataatgcagaccaggtcacccctgtttcccac attggctgccccaacactacacaa cagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgttgtgcaataca actctgaactggcacagccaagatg gagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaaaaggagttgg tggtagacagtcccgggctctact acgtatttttggaactgaagctcagtccaacattcacaaacacaggccacaaggtgcagg gctgggtctctcttgttttgcaagcaa agcctcaggtagatgactttgacaacttggccctgacagtggaactgttcccttgctcca tggagaacaagttagtggaccgttcct ggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtctgagggcttatc tgcatggagcccaggatgcataca gagactgggagctgtcttatcccaacaccaccagctttggactctttcttgtgaaacccg acaacccatgggaatgaGAATT

CAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGC

CGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCG

CTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGG

CGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCG

GTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCA

ACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTT

ACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGC

TTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAA

TCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCA

AAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGAC

GGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTT

CCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAG

GGATTTTGCCGATTTCGGTCT ATT GGTT AAA AAAT GAGCTGATTT AAC AAAA A

TTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCT

CAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCA

ACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAG

ACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

CACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT

TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAA

ATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC

CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA

GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT

TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG

AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG

TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT

TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAG

CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC AGT C AC AGAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGAATT AT GC AGT

GCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT

CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA

ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG

AGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT

AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG

AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG

GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTG

CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGAC

GGGGAGTC AGGC A ACT AT GGAT GA ACGA A AT AGAC AGATCGC T GAG AT AGG

TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATAC

TTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATC

CTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA

GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCT

GCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTT

TGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAG

CAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACC

ACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA

CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW102-pAAV EFSLTR-Cd80-sPA (SEQ ID NO: 36)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgTtgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgttagGAATT CAATAAAAGATCTT

TATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCCCTA

GTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGG

GCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGC

GAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTAC

GCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCC

CTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACC

GCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTT

CTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTT

AGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGG

GTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTG

ACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAAC

ACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTC

GGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTA

ACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCT

GATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGC

CCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTC

TCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGA

GACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATA

ATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCC

TATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATA

ACC CTGAT A A AT GC TT C A AT A AT ATTGA A A A AGGA AGAGT AT GAGT ATT C A A

CATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTG

CTC ACCC AGAAACGCTGGTGAAAGT AAAAGAT GCTGAAGATC AGTT GGGT GC

ACGAGTGGGTT AC ATCGAACTGGATCTC AAC AGCGGT AAGATCCTT GAGAGT

TTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATG

TGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGC

AT AC ACT ATTCTC AGAAT GACTT GGTTGAGT ACTC ACC AGT C AC AGAAA AGC

ATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCAT

GAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAG

GAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCG

TTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACG

AT GCCTGT AGC AAT GGC AAC AACGTTGCGC AAACT ATT AACTGGCGA ACT AC

TTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGT

TGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATA

AATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCC

AGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCA

AC T AT GG AT G A AC G A A AT AG AC AG AT C GC T GAG AT AGGT GC C TC AC T GATT A

AGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTA

AAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCT

CATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCG

TAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGC

TGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATC

AAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGAT

ACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACT

CTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCT

GCC AGT GGC GAT AAGTCGT GTCTT ACCGGGTT GGACTC AAGACGAT AGTT AC

CGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAG CTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGA

GAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGC

GGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCC

TGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTT

TTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGG

CCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW103-pAAV EFSLTR-Light-sPA (SEQ ID NO: 37)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggagagtgtggtacagccttcagtgtttgtggtggatggacagacggacatcccattc aggcggctggaacagaaccaccgg agacggcgctgtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctggg ctggccactcagggctggtttctc ctgagactgcatcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggc tcctgggagaagctgatacaagat caacgatctcaccaggccaacccagcagcacatcttacaggagccaacgccagcttgata ggtattggtggacctctgttatggg agacacgacttggcctggccttcttgaggggcttgacgtatcatgatggggccctggtga ccatggagcccggttactactatgtg tactccaaagtgcagctgagcggcgtgggctgcccccaggggctggccaatggcctcccc atcacccatggactatacaagcg cacatcccgctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccg ggccaacagctcccgagtctggt gggacagcagcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtcc gcgtgcctggaaaccgcctggt cagaccacgtgacggcaccaggtcctatttcggagctttcatggtctgaGAATTCAATAA AAGATCTTTATT

TTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCCCTAGTG

ATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCG

ACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAG

CGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCA

TCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTG

TAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCT

ACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTC

GCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGG

GTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTG

ATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACG

TTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACT

CAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGT

CTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACA

AAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGAT

GCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCT

GACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCC

GGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGAC

GAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATG

GTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTAT

TTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACC CTGAT A A AT GC TT C A AT A AT ATT GA A A A AGG A AGAGT AT GAGT ATT C A AC AT

TTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTC

ACCC AGA AACGCTGGT GAAAGT AAAAGATGCTGAAGAT C AGTTGGGTGC ACG

AGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTC

GCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGC

GCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATAC

ACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTT

ACGGATGGC AT GAC AGT AAGAGAATT ATGC AGTGCTGCC AT AACC ATGAGT G

ATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCT

AACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGG

AACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGC

CTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACT

CTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAG

GACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCT

GGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATG

GTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTAT

GGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCAT

TGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACT

TCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGA

CCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAA

AAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTG

CAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGC

TACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAAT

ACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGC

ACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG

GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAA

GGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAG

CGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCG

CCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGG

TCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCT

TTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATG

CTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTA

CGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW104-pAAV EFSLTR-Cxcl 10-sPA (SEQ ID NO: 38)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacccaagtgctgccgtcattttctgcctcatcctgctgggtctgagtgggactcaa gggatccctctcgcaaggacggtccg ctgcaactgcatccatatcgatgacgggccagtgagaatgagggccatagggaagcttga aatcatccctgcgagcctatcctg cccacgtgttgagatcattgccacgatgaaaaagaatgatgagcagagatgtctgaatcc ggaatctaagaccatcaagaatttaa tgaaagcgtttagccaaaaaaggtctaaaagggctccttgaGAATTCAATAAAAGATCTT TATTTTCAT

TAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCCCTAGTGATGGA

GTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAA

AGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGC

GCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGT

GCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGC

GGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACAC

TTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCA

CGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTC

CGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGG

TTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGG

AGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAAC

TCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTAT

T GGTT AAAAAAT GAGCTGATTT AAC AA AAATTT AACGCGAATTTT AAC AAAA

TATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCG

CATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACG

GGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGA

GCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAA

GGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTT

CTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGT

TTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTG

AT A A AT GCTT C A AT A AT ATT GA A A A AGG A AGAGT AT GAGT ATTC A AC ATTT C

CGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACC

C AG A A AC GC T GGT G A A AGT A A A AG AT GC T G A AG AT C AGT T GGGT GC AC GAGT

GGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCC

CCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCG

GTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACT

ATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACG

GAT GGC AT GAC AGT A AG AG A ATT AT GC AGT GCTGCC AT AAC CAT GAGT GAT A

ACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAAC

CGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAAC

CGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGT

AGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA

GCTTCCCGGC AAC AATT AAT AGACTGGAT GGAGGCGGAT AA AGTT GC AGGAC

CACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGA

GCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTA

AGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGA

TGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGG

TAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCA

TTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCA

AAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAG

ATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAA

ACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTAC

CAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACT

GTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC

GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCG

ATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGC

GCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGA

ACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCA CGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCG GAAC AGGAGAGCGC ACGAGGGAGCTTCC AGGGGGAAACGCCTGGT ATCTTT A TAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTC GTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW105-pAAV EFSLTR-41bbL-sPA (SEQ ID NO: 39)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggaccagcacacacttgatgtggaggataccgcggatgccagacatccagcaggtact tcgtgcccctcggatgcggcgct cctcagagataccgggctcctcgcggacgctgcgctcctctcagatactgtgcgccccac aaatgccgcgctccccacggatg ctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctgcactgaact tctgttcccgccacccaaagctctat ggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctatcttcacccgc accgagcctcggccagcgctcacaatca ccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacccctgtttccc acattggctgccccaacactacac aacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgttgtgcaata caactctgaactggcacagccaaga tggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaaaaggagtt ggtggtagacagtcccgggctcta ctacgtatttttggaactgaagctcagtccaacattcacaaacacaggccacaaggtgca gggctgggtctctcttgttttgcaagc aaagcctcaggtagatgactttgacaacttggccctgacagtggaactgttcccttgctc catggagaacaagttagtggaccgtt cctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtctgagggctt atctgcatggagcccaggatgcata cagagactgggagctgtcttatcccaacaccaccagctttggactctttcttgtgaaacc cgacaacccatgggaatgaGAAT

TCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGG

CCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTC

GCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGG

GCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGC

GGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGC

AACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGT

TACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCG

CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAA

ATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCC

AAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGA

CGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGT

TCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAA

GGGATTTTGCCGATTTCGGTCT ATT GGTT AAA AAAT GAGCTGATTT A AC AAAA

ATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTC

TCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCA

ACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAG

ACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

CACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT

TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAA ATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC

CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA

GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT

TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG

AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG

TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT

TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAG

CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC

AGT C AC AGAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGAATT AT GC AGT

GCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT

CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA

ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG

AGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT

AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG

AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG

GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTG

CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGAC

GGGGAGTC AGGC A ACT AT GGAT GA ACGA A AT AGAC AGATCGC T GAG AT AGG

TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATAC

TTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATC

CTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA

GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCT

GCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTT

TGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAG

CAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACC

ACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA

CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW 110-pAA V_EFS-LTR-Cd80-T2 A-Light-P2 A-Cxcl 10-E2 A-IFN g- WPRE (SEQ ID NO: 40)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccacc atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgctgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaAgtgctgggctggccactc agggctggtttctcctgagactgc atcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggaga agctgatacaagatcaacgatctc accaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtg gacctctgttatgggagacacgac ttggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagc ccggttactactatgtgtactccaaa gtgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccat ggactatacaagcgcacatccc gctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaaca gctcccgagtctggtgggacag cagcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcc tggaaaccgcctggtcagacca cgtgacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGC A AC AAACTTCTCTCT

GCTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgaacccaagtgctgc cg tcattttctgcctcatcctgctgggtctgagtgggactcaagggatccctctcgcaagga cggtccgctgcaactgcatccatatcg atgacgggccagtgagaatgagggccatagggaagcttgaaatcatccctgcgagcctat cctgcccacgtgttgagatcattg ccacgatgaaaaagaatgatgagcagagatgtctgaatccggaatctaagaccatcaaga atttaatgaaagcgtttagccaaaa aaggtctaaaagggctcctagcggtacccagtgcaccaactacgccctgctgaagctggc cggcgatgtggagagcaacccc gggcccgctagcatgaacgctacacactgcatcttggctttgcagctcttcctcatggct gtttctggctgttactgccacggcaca gtcattgaaagcctagaaagtctgaataactattttaactcaagtggcatagatgtggaa gaaaagagtctcttcttggatatctgga ggaactggcaaaaggatggtgacatgaaaatcctgcagagccagattatctctttctacc tcagactctttgaagtcttgaaagaca atcaggccatcagcaacaacataagcgtcattgaatcacacctgattactaccttcttca gcaacagcaaggcgaaaaaggatgc attcatgagtattgccaagtttgaggtcaacaacccacaggtgcagcgccaagcattcaa tgagctcatccgagtggtccaccag ctgttgccggaatccagcctcaggaagcggaaaaggagtcgctgctgaGaattcacgcgt taagtcgacaatcaacctctggat tacaaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgt ggatacgctgctttaatgcctttgtatcatgct attgcttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctt tatgaggagttgtggcccgttgtcaggcaacg tggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccac ctgtcagctcctttccgggactttcgc tttccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggac aggggctcggctgttgggcactgac aattccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgcc acctggattctgcgcgggacgtccttct gctacgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctc tgcggcctcttccgcgtcttcgccttc gccctcagacgagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaat gacGGCCGCAGGAACC

CCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGG

CCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGT

GAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTC

CTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTAC

GCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCG

TGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTT

CCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTC

CCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGA

TTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCC

CTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGA ACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCG

ATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAA

TTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCT

GCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGA

CGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGA

CCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGC

GCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGAT

AATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGA

ACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGA

C A AT A AC CCTGAT A A AT GC TT C A AT A AT ATT GA A A A AGGA AGAGT AT GAGT A

TTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGT

TTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTG

GGT GC AC GAGT GGGTT AC ATCGA ACTGGAT C TC A AC AGC GGT A AG AT C C TTG

AGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTG

CTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTC

GCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAA

AAGC ATCTT ACGGAT GGC AT GAC AGT AAGAGAATT AT GC AGT GCTGCC AT AA

CCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACC

GAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTG

ATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACAC

C AC GAT GCCTGT AGC AAT GGC AAC AACGTT GCGC A AACT ATT AACTGGCGAA

CTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATA

AAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCT

GATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGG

GGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCA

GGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTG

ATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGA

TTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATA

ATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGAC

CCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAAT

CTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGG

ATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCA

GATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGA

ACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCT

GCTGCC AGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTT

ACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCC

AGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTAT

GAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAA

GCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACG

CCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGAT

TTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGC

GGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW 111-pAA V_EFS-LTR-Light-P2 A-Cxcl 10-E2 A-IFN g- WPRE (SEQ ID NO: 41)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggagagtgtggtacagccttcagtgtttgtggtggatggacagacggacatcccattc aggcggctggaacagaaccaccgg agacggcgctgtggcactgtccaggtcagcctggccctggtgctgctgctaAgtgctggg ctggccactcagggctggtttctc ctgagactgcatcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggc tcctgggagaagctgatacaagat caacgatctcaccaggccaacccagcagcacatcttacaggagccaacgccagcttgata ggtattggtggacctctgttatggg agacacgacttggcctggccttcttgaggggcttgacgtatcatgatggggccctggtga ccatggagcccggttactactatgtg tactccaaagtgcagctgagcggcgtgggctgcccccaggggctggccaatggcctcccc atcacccatggactatacaagcg cacatcccgctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccg ggccaacagctcccgagtctggt gggacagcagcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtcc gcgtgcctggaaaccgcctggt cagaccacgtgacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGG CGCAACAAACTTC

TCTCTGCTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatgaacccaagt gctgccgtcattttctgcctcatcctgctgggtctgagtgggactcaagggatccctctc gcaaggacggtccgctgcaactgcat ccatatcgatgacgggccagtgagaatgagggccatagggaagcttgaaatcatccctgc gagcctatcctgcccacgtgttga gatcattgccacgatgaaaaagaatgatgagcagagatgtctgaatccggaatctaagac catcaagaatttaatgaaagcgttta gccaaaaaaggtctaaaagggctcctagcggtacccagtgcaccaactacgccctgctga agctggccggcgatgtggagag caaccccgggcccgctagcatgaacgctacacactgcatcttggctttgcagctcttcct catggctgtttctggctgttactgcca cggcacagtcattgaaagcctagaaagtctgaataactattttaactcaagtggcataga tgtggaagaaaagagtctcttcttgga tatctggaggaactggcaaaaggatggtgacatgaaaatcctgcagagccagattatctc tttctacctcagactctttgaagtcttg aaagacaatcaggccatcagcaacaacataagcgtcattgaatcacacctgattactacc ttcttcagcaacagcaaggcgaaaa aggatgcattcatgagtattgccaagtttgaggtcaacaacccacaggtccagcgccaag cattcaatgagctcatccgagtggt ccaccagctgttgccggaatccagcctcaggaagcggaaaaggagtcgctgctgaGaatt cacgcgttaagtcgacaatcaac ctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgctcctttta cgctatgtggatacgctgctttaatgcctttgt atcatgctattgcttcccgtatggctttcattttctcctccttgtataaatcctggttgc tgtctctttatgaggagttgtggcccgttgtca ggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattg ccaccacctgtcagctcctttccgg gactttcgctttccccctccctattgccacggcggaactcatcgccgcctgccttgcccg ctgctggacaggggctcggctgttgg gcactgacaattccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcct gtgttgccacctggattctgcgcggga cgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcccgcggcctgc tgccggctctgcggcctcttccgcgtc ttcgccttcgccctcagacgagtcggatctccctttgggccgcctccccgcgtcgacttt aagaccaatgacGGCCGCAG

GAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCAC

TGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCC

TCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATT

TTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCAT

AGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCG

CAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCT

TCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGG

GGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAA

CTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTT

TCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAAC

TGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTT

GCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACG

CGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACA

ATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCG

CTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCT

GTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAA ACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCA

TGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGC

GGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATG

AGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGA

GTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCC

TGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAG

TTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCC

TTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTT

CTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCG

GTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACA

GAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGA ATT ATGC AGTGCTGCC A

TAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGG

ACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGC

CTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTG

ACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGG

CGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCG

GATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTAT

TGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCA

CTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGA

GT C AGGC AACT ATGGAT GAACGAAAT AGAC AGATCGCTGAGAT AGGT GCCTC

ACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGA

TTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTG

ATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCA

GACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGT

AATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGC

CGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGC

GCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCA

AGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTG

GCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGAT

AGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACA

GCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAG

CTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCG

GT AAGCGGC AGGGTCGGAAC AGGAGAGCGC ACGAGGGAGCTTCC AGGGGGA

AACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCG

TCGATTTTT GTGAT GCTCGT C AGGGGGGCGGAGCCT AT GGAAAAACGCC AGC

AACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW 112-pAA V EFS-LTR-41BBL-T2 A-Light-P2 A-Cxcl 10-E2 A-IFN g- WPRE (SEQ ID NO: 42)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggaccagcacacacttgatgtggaggataccgcggatgccagacatccagcaggtact tcgtgcccctcggatgcggcgct cctcagagataccgggctcctcgcggacgctgcgctcctctcagatactgtgcgccccac aaatgccgcgctccccacggatg ctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctgcactgaact tctgttcccgccacccaaagctctat ggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctatcttcacccgc accgagcctcggccagcgctcacaatca ccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacccctgtttccc acattggctgccccaacactacac aacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgttgtgcaata caactctgaactggcacagccaaga tggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaaaaggagtt ggtggtagacagtcccgggctcta ctacgtatttttggaactgaagctcagtccaacattcacaaacacaggccacaaggtgca gggctgggtctctcttgttttgcaagc aaagcctcaggtagatgactttgacaacttggccctgacagtggaactgttcccttgctc catggagaacaagttagtggaccgtt cctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtctgagggctt atctgcatggagcccaggatgcata cagagactgggagctgtcttatcccaacaccaccagctttggactctttcttgtgaaacc cgacaacccatgggaaGGCAGT

GGAGAGGGCAGAGGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCT

GGCCCAatggagagtgtggtacagccttcagtgtttgtggtggatggacagacggac atcccattcaggcggctggaaca gaaccaccggagacggcgctgtggcactgtccaggtcagcctggccctggtgctgctgct aAgtgctgggctggccactcag ggctggtttctcctgagactgcatcaacgtcttggagacatagtagctcatctgccagat ggaggcaaaggctcctgggagaagc tgatacaagatcaacgatctcaccaggccaacccagcagcacatcttacaggagccaacg ccagcttgataggtattggtggac ctctgttatgggagacacgacttggcctggccttcttgaggggcttgacgtatcatgatg gggccctggtgaccatggagcccgg ttactactatgtgtactccaaagtgcagctgagcggcgtgggctgcccccaggggctggc caatggcctccccatcacccatgg actatacaagcgcacatcccgctacccgaaggagttagaactgctggtcagtcggcggtc accctgtggccgggccaacagct cccgagtctggtgggacagcagcttcctgggcggcgtggtacatctggaggctggggaag aggtggtggtccgcgtgcctgg aaaccgcctggtcagaccacgtgacggcaccaggtcctatttcggagctttcatggtcac tagtGGATCCGGCGCAA

CAAACTTCTCTCTGCTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACC

Gatgaacccaagtgctgccgtcattttctgcctcatcctgctgggtctgagtgggac tcaagggatccctctcgcaaggacggtc cgctgcaactgcatccatatcgatgacgggccagtgagaatgagggccatagggaagctt gaaatcatccctgcgagcctatcc tgcccacgtgttgagatcattgccacgatgaaaaagaatgatgagcagagatgtctgaat ccggaatctaagaccatcaagaattt aatgaaagcgtttagccaaaaaaggtctaaaagggctcctagcggtacccagtgcaccaa ctacgccctgctgaagctggccg gcgatgtggagagcaaccccgggcccgctagcatgaacgctacacactgcatcttggctt tgcagctcttcctcatggctgtttct ggctgttactgccacggcacagtcattgaaagcctagaaagtctgaataactattttaac tcaagtggcatagatgtggaagaaaa gagtctcttcttggatatctggaggaactggcaaaaggatggtgacatgaaaatcctgca gagccagattatctctttctacctcag actctttgaagtcttgaaagacaatcaggccatcagcaacaacataagcgtcattgaatc acacctgattactaccttcttcagcaac agcaaggcgaaaaaggatgcattcatgagtattgccaagtttgaggtcaacaacccacag gtccagcgccaagcattcaatgag ctcatccgagtggtccaccagctgttgccggaatccagcctcaggaagcggaaaaggagt cgctgctgaGaattcacgcgtta agtcgacaatcaacctctggattacaaaatttgtgaaagattgactggtattcttaacta tgttgctccttttacgctatgtggatacgct gctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttg tataaatcctggttgctgtctctttatgaggagtt gtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccac tggttggggcattgccaccacctgt cagctcctttccgggactttcgctttccccctccctattgccacggcggaactcatcgcc gcctgccttgcccgctgctggacagg ggctcggctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttcc ttggctgctcgcctgtgttgccacctg gattctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttcc ttcccgcggcctgctgccggctctgc ggcctcttccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcct ccccgcgtcgactttaagaccaatga cGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCG

CTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCC

CGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGA

TGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAA

AGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGT

GGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTT

TCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTC

TAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGAC

CCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATA GACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTT

GTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTAT

AAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAA

AAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCAC

TCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGC

CAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTAC

AGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTC

ATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAG

GTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGG

AAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGT

ATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAG

GAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGC

ATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATG

CTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAG

CGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCA

CTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAA

GAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTC

ACC AGT C AC AGAAAAGC ATCTT ACGGAT GGC AT GAC AGT AAGAGAATT AT GC

AGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAAC

GATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCAT

GTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACG

ACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACT

ATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGA

TGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGG

CTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATC

ATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACAC

GACGGGGAGT C AGGC AACT ATGGAT GAACGAAAT AGAC AGATCGCTGAGAT

AGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATA

TACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAG

ATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCAC

TGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTT

TCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTG

GTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTT

CAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCC

ACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTG

TTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC

AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCG

TGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTAC

AGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA

GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTC

CAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGA

CTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAA

ACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTC

ACATGT pGW 113-pAA V EFS-LTR-IL23-T2 A-Light-P2 A-Cxcl 10-E2 A-IFN g- WPRE (SEQ ID NO: 43)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGCTGGTGTCT

CCACTCATGGCCATGTGGGAGCTGGAGAAAGACGTTTATGTTGTAGAGGTGG

ACTGGACTCCCGATGCCCCTGGAGAAACAGTGAACCTCACCTGTGACACGCC

TGAAGAAGATGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGC

TCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTTCTAGATGCTGGCCAGTA

CACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCAC

AAGAAGGAAAAT GGAATTT GGTCC ACTGAAATTTT AAAAAATTT C AAAA AC A

AGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCA

T GGCTGGTGC A AAGAAAC ATGGACTTGAAGTTC AAC AT C AAGAGC AGT AGC A

GTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAG

AAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGG

AGGATGTCACCTGCCCAACTGCCGAGGAGACCCTGCCCATTGAACTGGCGTT

GGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATC

AGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGA

AGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCTGACTCCTGGAGCACTCC

CCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAAA

AG AT GA AGGAGAC AGAGGAGGGGT GT AAC C AGA A AGGT GC GTTCCTCGT AG

AGAAGAC ATCT ACCGAAGTCC AATGC AAAGGCGGGAAT GTCTGCGT GC AAGC

TCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCA

GGGTCCGATCCGGCAGTGGAGAGGGCAGAGGAAGTCTGCTAACATGCGGTGA

CGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgtttgtggt ggatggacagacgg acatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcactgtccagg tcagcctggccctggtgctgctg ctaAgtgctgggctggccactcagggctggtttctcctgagactgcatcaacgtcttgga gacatagtagctcatctgccagatg gaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggccaacccag cagcacatcttacaggagccaa cgccagcttgataggtattggtggacctctgttatgggagacacgacttggcctggcctt cttgaggggcttgacgtatcatgatgg ggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagctgagcgg cgtgggctgcccccaggggctgg ccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccgaaggagt tagaactgctggtcagtcggcggt caccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgggcggcg tggtacatctggaggctgggga agaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcaccaggtc ctatttcggagctttcatggtcact agtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATGTCGA

AGAGAATCCTGGACCGatgaacccaagtgctgccgtcattttctgcctcatcctgct gggtctgagtgggactcaa gggatccctctcgcaaggacggtccgctgcaactgcatccatatcgatgacgggccagtg agaatgagggccatagggaagct tgaaatcatccctgcgagcctatcctgcccacgtgttgagatcattgccacgatgaaaaa gaatgatgagcagagatgtctgaatc cggaatctaagaccatcaagaatttaatgaaagcgtttagccaaaaaaggtctaaaaggg ctcctagcggtacccagtgcaccaa ctacgccctgctgaagctggccggcgatgtggagagcaaccccgggcccgctagcatgaa cgctacacactgcatcttggcttt gcagctcttcctcatggctgtttctggctgttactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaa gtggcatagatgtggaagaaaagagtctcttcttggatatctggaggaactggcaaaagg atggtgacatgaaaatcctgcagag ccagattatctctttctacctcagactctttgaagtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacac ctgattactaccttcttcagcaacagcaaggcgaaaaaggatgcattcatgagtattgcc aagtttgaggtcaacaacccacaggt ccagcgccaagcattcaatgagctcatccgagtggtccaccagctgttgccggaatccag cctcaggaagcggaaaaggagtc gctgctgaGaattcacgcgttaagtcgacaatcaacctctggattacaaaatttgtgaaa gattgactggtattcttaactatgttgct ccttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttcccgt atggctttcattttctcctccttgtataaatcct ggttgctgtctctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtgtgca ctgtgtttgctgacgcaacccccactg gttggggcattgccaccacctgtcagctcctttccgggactttcgctttccccctcccta ttgccacggcggaactcatcgccgcct gccttgcccgctgctggacaggggctcggctgttgggcactgacaattccgtggtgttgt cggggaaatcatcgtcctttccttgg ctgctcgcctgtgttgccacctggattctgcgcgggacgtccttctgctacgtcccttcg gccctcaatccagcggaccttccttcc cgcggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagacgagt cggatctccctttgggccgcctccc cgcgtcgactttaagaccaatgacGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTC C

CTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGA

CGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCC

TGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCA

CACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAG

CGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCC

TTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGC

TTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGC

TTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTG

GGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTC

TTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGG

CTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAA

TGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTA

CAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGC

CAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGC

TCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGT

CAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGA

TACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCA

GGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTA

AATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTC

AATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTT

ATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTG

GTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCG

AACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACG

TTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCC

GTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAA

TGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATG

ACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGG

CCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTG

CACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGA

ATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGC

AACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGC

AACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG

CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGC

GTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCG

T ATC GT AGT TATCTACAC G AC GGGG AGT C AGGC A AC TAT GG AT G A AC G A A AT

AGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAG

ACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTA

AAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAA

CGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGAT

CTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA

CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTT

TCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAG TGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATAC

CTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTG

TCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCG

GGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACA

CCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGA

AGGGAGA A AGGC GGAC AGGT ATCCGGT A AGC GGC AGGGT C GGA AC AGGAGA

GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTC

GGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGG

GCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCC

TTTTGCTGGCCTTTTGCTCACATGT pGW 114-pAA V_EFS-LTR-Cd80-T2 A-Light-P2 A-41BBL-E2 A-IFN g- WPRE (SEQ ID NO: 44)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgctgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttccttcgtacgGGCAG TGGAGAGGGCAGA

GGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgt ggtacagccttcagtgtttgtggtggatggacagacggacatcccattcaggcggctgga acagaaccaccggagacggcgct gtggcactgtccaggtcagcctggccctggtgctgctgctaAgtgctgggctggccactc agggctggtttctcctgagactgc atcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggctcctgggaga agctgatacaagatcaacgatctc accaggccaacccagcagcacatcttacaggagccaacgccagcttgataggtattggtg gacctctgttatgggagacacgac ttggcctggccttcttgaggggcttgacgtatcatgatggggccctggtgaccatggagc ccggttactactatgtgtactccaaa gtgcagctgagcggcgtgggctgcccccaggggctggccaatggcctccccatcacccat ggactatacaagcgcacatccc gctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccgggccaaca gctcccgagtctggtgggacag cagcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtccgcgtgcc tggaaaccgcctggtcagacca cgtgacggcaccaggtcctatttcggagctttcatggtcactagtGGATCCGGCGC A AC AAACTTCTCTCT

GCTGA A AC A AGCC GGAGAT GT C GA AGAGA ATCC T GGAC CGatggaccagcacacacttg atgtggaggataccgcggatgccagacatccagcaggtacttcgtgcccctcggatgcgg cgctcctcagagataccgggctc ctcgcggacgctgcgctcctctcagatactgtgcgccccacaaatgccgcgctccccacg gatgctgcctaccctgcggttaatg ttcgggatcgcgaggccgcgtggccgcctgcactgaacttctgttcccgccacccaaagc tctatggcctagtcgctttggttttg ctgcttctgatcgccgcctgtgttcctatcttcacccgcaccgagcctcggccagcgctc acaatcaccacctcgcccaacctgg gtacccgagagaataatgcagaccaggtcacccctgtttcccacattggctgccccaaca ctacacaacagggctctcctgtgtt cgccaagctactggctaaaaaccaagcatcgttgtgcaatacaactctgaactggcacag ccaagatggagctgggagctcata cctatctcaaggtctgaggtacgaagaagacaaaaaggagttggtggtagacagtcccgg gctctactacgtatttttggaactga agctcagtccaacattcacaaacacaggccacaaggtgcagggctgggtctctcttgttt tgcaagcaaagcctcaggtagatga ctttgacaacttggccctgacagtggaactgttcccttgctccatggagaacaagttagt ggaccgttcctggagtcaactgttgct cctgaaggctggccaccgcctcagtgtgggtctgagggcttatctgcatggagcccagga tgcatacagagactgggagctgtc ttatcccaacaccaccagctttggactctttcttgtgaaacccgacaacccatgggaaag cggtacccagtgcaccaactacgcc ctgctgaagctggccggcgatgtggagagcaaccccgggcccgctagcatgaacgctaca cactgcatcttggctttgcagctc ttcctcatggctgtttctggctgttactgccacggcacagtcattgaaagcctagaaagt ctgaataactattttaactcaagtggcat agatgtggaagaaaagagtctcttcttggatatctggaggaactggcaaaaggatggtga catgaaaatcctgcagagccagatt atctctttctacctcagactctttgaagtcttgaaagacaatcaggccatcagcaacaac ataagcgtcattgaatcacacctgatta ctaccttcttcagcaacagcaaggcgaaaaaggatgcattcatgagtattgccaagtttg aggtcaacaacccacaggtgcagcg ccaagcattcaatgagctcatccgagtggtccaccagctgttgccggaatccagcctcag gaagcggaaaaggagtcgctgct gaGaattcacgcgttaagtcgacaatcaacctctggattacaaaatttgtgaaagattga ctggtattcttaactatgttgctcctttta cgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttcccgtatggctt tcattttctcctccttgtataaatcctggttgct gtctctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtt tgctgacgcaacccccactggttggg gcattgccaccacctgtcagctcctttccgggactttcgctttccccctccctattgcca cggcggaactcatcgccgcctgccttg cccgctgctggacaggggctcggctgttgggcactgacaattccgtggtgttgtcgggga aatcatcgtcctttccttggctgctc gcctgtgttgccacctggattctgcgcgggacgtccttctgctacgtcccttcggccctc aatccagcggaccttccttcccgcgg cctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagacgagtcggat ctccctttgggccgcctccccgcgtc gactttaagaccaatgacGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTC

TGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCC

GGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAG

GGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCG

CATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGG

CGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGC

GCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCC

CGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACG

GCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCAT

CGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATA

GTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCT

TTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCT

GATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTT

TATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCC

CGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGG

CATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAG

GTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGC

CTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGG

CACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATAC

ATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAA

TATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCC

_{CTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAA}

AGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTG

GATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCC

AATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTG

ACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTT

GGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTA

AGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACT TACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAA

CATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAA

GCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAA

CGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAA

TT AAT AGACTGGAT GGAGGCGGAT AAAGTT GC AGGACC ACTTCTGCGCTCGG

CCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGA

AGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCG

T AGTT ATCTACAC G AC GGGG AGT C AGGC A AC TAT GG AT G A AC G A A AT AG AC A

GATCGCTGAGAT AGGT GCCTC ACTGATT AAGC ATTGGT AACTGT C AGACC AA

GTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGG

ATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGA

GTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTT

GAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCG

CTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAA

GGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGC

CGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCT

CTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTAC

CGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGA

ACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAAC

TGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAG

AAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCAC

GAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTC

GCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGC

CTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTG

GCCTTTTGCTCACATGT pGW 115-pAAV_EFS-LTR-IL23-T2A-Light-P2A-41BBL-E2A-IFNg-WPRE (SEQ ID NO: 45)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGCTGGTGTCT

CCACTCATGGCCATGTGGGAGCTGGAGAAAGACGTTTATGTTGTAGAGGTGG

ACTGGACTCCCGATGCCCCTGGAGAAACAGTGAACCTCACCTGTGACACGCC

TGAAGAAGATGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGC

TCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTTCTAGATGCTGGCCAGTA

CACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCAC

AAGAAGGAAAAT GGAATTTGGTCC ACTGAAATTTT AAAAAATTT C AAAA AC A

AGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCA

TGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCA GTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAG

AAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGG

AGGATGTCACCTGCCCAACTGCCGAGGAGACCCTGCCCATTGAACTGGCGTT

GGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATC

AGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGA

AGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCTGACTCCTGGAGCACTCC

CCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAAA

AG AT G A AGG AG AC AG AGG AGGGGT GT A AC C AG A A AGGT GC GT TC C T C GT AG

AGAAGAC ATCT ACCGAAGTCC AATGC AAAGGCGGGAAT GTCTGCGT GC AAGC

TCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCA

GGGTCCGATCCGGCAGTGGAGAGGGCAGAGGAAGTCTGCTAACATGCGGTGA

CGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgtttgtggt ggatggacagacgg acatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcactgtccagg tcagcctggccctggtgctgctg ctaAgtgctgggctggccactcagggctggtttctcctgagactgcatcaacgtcttgga gacatagtagctcatctgccagatg gaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggccaacccag cagcacatcttacaggagccaa cgccagcttgataggtattggtggacctctgttatgggagacacgacttggcctggcctt cttgaggggcttgacgtatcatgatgg ggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagctgagcgg cgtgggctgcccccaggggctgg ccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccgaaggagt tagaactgctggtcagtcggcggt caccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgggcggcg tggtacatctggaggctgggga agaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcaccaggtc ctatttcggagctttcatggtcact agtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATGTCGA

AGAGAATCCTGGACCGatggaccagcacacacttgatgtggaggataccgcggatgc cagacatccagcaggt acttcgtgcccctcggatgcggcgctcctcagagataccgggctcctcgcggacgctgcg ctcctctcagatactgtgcgcccc acaaatgccgcgctccccacggatgctgcctaccctgcggttaatgttcgggatcgcgag gccgcgtggccgcctgcactgaa cttctgttcccgccacccaaagctctatggcctagtcgctttggttttgctgcttctgat cgccgcctgtgttcctatcttcacccgcac cgagcctcggccagcgctcacaatcaccacctcgcccaacctgggtacccgagagaataa tgcagaccaggtcacccctgttt cccacattggctgccccaacactacacaacagggctctcctgtgttcgccaagctactgg ctaaaaaccaagcatcgttgtgcaat acaactctgaactggcacagccaagatggagctgggagctcatacctatctcaaggtctg aggtacgaagaagacaaaaagga gttggtggtagacagtcccgggctctactacgtatttttggaactgaagctcagtccaac attcacaaacacaggccacaaggtgc agggctgggtctctcttgttttgcaagcaaagcctcaggtagatgactttgacaacttgg ccctgacagtggaactgttcccttgctc catggagaacaagttagtggaccgttcctggagtcaactgttgctcctgaaggctggcca ccgcctcagtgtgggtctgagggct tatctgcatggagcccaggatgcatacagagactgggagctgtcttatcccaacaccacc agctttggactctttcttgtgaaaccc gacaacccatgggaaagcggtacccagtgcaccaactacgccctgctgaagctggccggc gatgtggagagcaaccccggg cccgctagcatgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtt tctggctgttactgccacggcacagtca ttgaaagcctagaaagtctgaataactattttaactcaagtggcatagatgtggaagaaa agagtctcttcttggatatctggaggaa ctggcaaaaggatggtgacatgaaaatcctgcagagccagattatctctttctacctcag actctttgaagtcttgaaagacaatca ggccatcagcaacaacataagcgtcattgaatcacacctgattactaccttcttcagcaa cagcaaggcgaaaaaggatgcattc atgagtattgccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgag ctcatccgagtggtccaccagctgt tgccggaatccagcctcaggaagcggaaaaggagtcgctgctgaGaattcacgcgttaag tcgacaatcaacctctggattaca aaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgtggat acgctgctttaatgcctttgtatcatgctattg cttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctttatg aggagttgtggcccgttgtcaggcaacgtgg cgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccacctg tcagctcctttccgggactttcgctttc cccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggacaggg gctcggctgttgggcactgacaatt ccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgccacct ggattctgcgcgggacgtccttctgcta cgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctctgcg gcctcttccgcgtcttcgccttcgcc ctcagacgagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaatgac GGCCGCAGGAACCCC

TAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCC

GGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGA

GCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTT

ACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGC GCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTG

ACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCC

TTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCC

TTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATT

TGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCT

TTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAAC

AACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGAT

TTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATT

TTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGC

TCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACG

CGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACC

GTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGC

GAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAA

TAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACC

CCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAA

T A ACCC T GAT A A AT GCTT C A AT A AT ATT GA A A A AGGA AGAGT AT GAGT ATTC

AACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTT

T GCTC ACCC AGAAACGCTGGTGAA AGT A AAAGAT GCTGAAGAT C AGTTGGGT

GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGA

GTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTA

TGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCC

GCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA

GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACC

ATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA

AGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGAT

CGTTGGGAACCGGAGCTGAAT GAAGCC AT ACC AAACGACGAGCGT GAC ACC A

CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACT

ACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAA

GTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGA

T A A AT C T GG AGC C GGT G AGC GT GG A AGC C GC GGT AT C ATT GC AGC AC T GGGG

CCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG

CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGAT

TAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATT

TAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAAT

CTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCC

CGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCT

GCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGA

TCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAG

ATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA

CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTG

CTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTA

CCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCA

GCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATG

AGAAAGCGCC ACGCTTCCCGAAGGGAGAAAGGCGGAC AGGT ATCCGGT AAG

CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGC

CTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATT

TTTGT GAT GCTCGT C AGGGGGGCGGAGCCT AT GGAAAAACGCC AGC AACGCG

GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW118-pAAV EFSLTR-Ifng-P2A-Cd80-T2A-light-E2A-41BBL-sPA (SEQ ID NO: 46)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGC AGTGGAGAGGGC AGAGGAAGTCTGCT AAC AT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctgc cagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcca acccagcagcacatcttacagg agccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcct ggccttcttgaggggcttgacgtatc atgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagc tgagcggcgtgggctgcccccagg ggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccga aggagttagaactgctggtcagtc ggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgg gcggcgtggtacatctggaggc tggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcac caggtcctatttcggagctttcat ggtcactagcagcggtacccagtgcaccaactacgccctgctgaagctggccggcgatgt ggagagcaaccccgggcccatg gaccagcacacacttgatgtggaggataccgcggatgccagacatccagcaggtacttcg tgcccctcggatgcggcgctcct cagagataccgggctcctcgcggacgctgcgctcctctcagatactgtgcgccccacaaa tgccgcgctccccacggatgctg cctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctgcactgaacttct gttcccgccacccaaagctctatgg cctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctatcttcacccgcac cgagcctcggccagcgctcacaatcacc acctcgcccaacctgggtacccgagagaataatgcagaccaggtcacccctgtttcccac attggctgccccaacactacacaa cagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgttgtgcaataca actctgaactggcacagccaagatg gagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaaaaggagttgg tggtagacagtcccgggctctact acgtatttttggaactgaagctcagtccaacattcacaaacacaggccacaaggtgcagg gctgggtctctcttgttttgcaagcaa agcctcaggtagatgactttgacaacttggccctgacagtggaactgttcccttgctcca tggagaacaagttagtggaccgttcct ggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtctgagggcttatc tgcatggagcccaggatgcataca gagactgggagctgtcttatcccaacaccaccagctttggactctttcttgtgaaacccg acaacccatgggaatgaGAATT

CAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGC

CGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCG

CTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGG

CGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCG

GTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCA

ACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTT

ACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGC

TTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAA

TCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCA

AAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGAC

GGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTT

CCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAG

GGATTTTGCCGATTTCGGTCT ATT GGTT AAA AAAT GAGCTGATTT AAC AAAA A

TTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCT

CAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCA

ACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAG

ACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

CACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT

TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAA

ATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC

CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA

GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT

TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG

AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG

TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT

TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAG

CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC

AGT C AC AGAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGAATT AT GC AGT

GCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT

CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA

ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG

AGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT

AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG

AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG

GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTG

CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGAC

GGGGAGTC AGGC A ACT AT GGAT GA ACGA A AT AGAC AGATCGC T GAG AT AGG

TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATAC

TTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATC

CTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA

GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCT

GCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTT

TGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAG

CAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACC

ACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA

CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW119-pAAV EFSLTR-Ifng-STOP-P2A-Cd80-T2A-light-P2A-E2A-41BBL-sPA (SEQ ID NO: 47)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGT AT AT AAGT GC AGT AGTCGCCGT GAACGTTCTTTTTCGC AAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcTGAATCCGGCGCAACAAACTTCTCTCTG CTGA

AACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttga tgcagg atacaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattc gtctttcacaagtgtcttcagatgttgatgaa caactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctac tggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggccc gagtataagaaccggactttatatg acaacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaaga ggaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctct acccccaacataactgagtctggaaac ccatctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaa ttacctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagt agccaactagatttcaatacgactcgca accacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcct gatagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtc gtcatcgttgtcatcatcaaatgcttct gtaagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagcctta ccttcgggcctgaagaagcatta gctgaacagaccgtcttccttcgtacgGGCAGTGGAGAGGGCAGAGGAAGTCTGCTAACA TG

CGGT GAC GT C GAGGAGA ATCC T GGC CC Aatggagagtgtggtacagccttcagtgtttgtggtggatg gacagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggca ctgtccaggtcagcctggccct ggtgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatca acgtcttggagacatagtagctcatc tgccagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccagg ccaacccagcagcacatcttaca ggagccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggc ctggccttcttgaggggcttgacgt atcatgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgc agctgagcggcgtgggctgccccc aggggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacc cgaaggagttagaactgctggtc agtcggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttc ctgggcggcgtggtacatctgga ggctggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacgg caccaggtcctatttcggagctt tcatggtcactagcagcggtacccagtgcaccaactacgccctgctgaagctggccggcg atgtggagagcaaccccgggcc catggaccagcacacacttgatgtggaggataccgcggatgccagacatccagcaggtac ttcgtgcccctcggatgcggcgc tcctcagagataccgggctcctcgcggacgctgcgctcctctcagatactgtgcgcccca caaatgccgcgctccccacggatg ctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctgcactgaact tctgttcccgccacccaaagctctat ggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctatcttcacccgc accgagcctcggccagcgctcacaatca ccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacccctgtttccc acattggctgccccaacactacac aacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgttgtgcaata caactctgaactggcacagccaaga tggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaaaaggagtt ggtggtagacagtcccgggctcta ctacgtatttttggaactgaagctcagtccaacattcacaaacacaggccacaaggtgca gggctgggtctctcttgttttgcaagc aaagcctcaggtagatgactttgacaacttggccctgacagtggaactgttcccttgctc catggagaacaagttagtggaccgtt cctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtctgagggctt atctgcatggagcccaggatgcata cagagactgggagctgtcttatcccaacaccaccagctttggactctttcttgtgaaacc cgacaacccatgggaatgaGAAT

TCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGG

CCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTC

GCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGG

GCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGC

GGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGC

AACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGT

TACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCG

CTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAA

ATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCC

AAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGA

CGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGT

TCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAA

GGGATTTTGCCGATTTCGGTCT ATT GGTT AAA AAAT GAGCTGATTT A AC AAAA

ATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTC

TCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCA

ACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAG

ACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

CACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT

TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAA

ATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC

CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA

GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT

TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG

AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG

TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT

TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAG

CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC

AGT C AC AGAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGAATT AT GC AGT

GCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT

CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA

ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG

AGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT

AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG

AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG

GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTG

CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGAC

GGGGAGTC AGGC A ACT AT GGAT GA ACGA A AT AGAC AGATCGC T GAG AT AGG TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATAC

TTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATC

CTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA

GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCT

GCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTT

TGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAG

CAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACC

ACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA

CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW122-pAAV EFS-LTR-IL23-T2A-41BBL-P2A-IFNg-WPRE (SEQ ID NO: 48)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGCTGGTGTCT

CCACTCATGGCCATGTGGGAGCTGGAGAAAGACGTTTATGTTGTAGAGGTGG

ACTGGACTCCCGATGCCCCTGGAGAAACAGTGAACCTCACCTGTGACACGCC

TGAAGAAGATGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGC

TCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTTCTAGATGCTGGCCAGTA

CACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCAC

AAGAAGGAAAAT GGAATTT GGTCC ACTGAAATTTT AAAAAATTT C AAAA AC A

AGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCA

T GGCTGGTGC A AAGAAAC ATGGACTTGAAGTTC AAC AT C AAGAGC AGT AGC A

GTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAG

AAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGG

AGGATGTCACCTGCCCAACTGCCGAGGAGACCCTGCCCATTGAACTGGCGTT

GGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATC

AGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGA

AGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCTGACTCCTGGAGCACTCC

CCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAAA

AG AT G A AGG AG AC AG AGG AGGGGT GT AAC C AG A A AGGT GC GT TC C T C GT AG

AGAAGAC ATCT ACCGAAGTCC AATGC AAAGGCGGGAAT GTCTGCGT GC AAGC TCAGGATCGCTATTACAATTCCTCATGCAGCAAGTGGGCATGTGTTCCCTGCA

GGGTCCGATCCGGCAGTGGAGAGGGCAGAGGAAGTCTGCTAACATGCGGTGA

CGTCGAGGAGAATCCTGGCCCAatggaccagcacacacttgatgtggaggataccgc ggatgccagacat ccagcaggtacttcgtgcccctcggatgcggcgctcctcagagataccgggctcctcgcg gacgctgcgctcctctcagatact gtgcgccccacaaatgccgcgctccccacggatgctgcctaccctgcggttaatgttcgg gatcgcgaggccgcgtggccgcc tgcactgaacttctgttcccgccacccaaagctctatggcctagtcgctttggttttgct gcttctgatcgccgcctgtgttcctatcttc acccgcaccgagcctcggccagcgctcacaatcaccacctcgcccaacctgggtacccga gagaataatgcagaccaggtca cccctgtttcccacattggctgccccaacactacacaacagggctctcctgtgttcgcca agctactggctaaaaaccaagcatcg ttgtgcaatacaactctgaactggcacagccaagatggagctgggagctcatacctatct caaggtctgaggtacgaagaagac aaaaaggagttggtggtagacagtcccgggctctactacgtatttttggaactgaagctc agtccaacattcacaaacacaggcc acaaggtgcagggctgggtctctcttgttttgcaagcaaagcctcaggtagatgactttg acaacttggccctgacagtggaactg ttcccttgctccatggagaacaagttagtggaccgttcctggagtcaactgttgctcctg aaggctggccaccgcctcagtgtggg tctgagggcttatctgcatggagcccaggatgcatacagagactgggagctgtcttatcc caacaccaccagctttggactctttct tgtgaaacccgacaacccatgggaaGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACA A

GCCGGAGATGTCGAAGAGAATCCTGGACCGgctagcatgaacgctacacactgcatc ttggctttgc agctcttcctcatggctgtttctggctgttactgccacggcacagtcattgaaagcctag aaagtctgaataactattttaactcaagt ggcatagatgtggaagaaaagagtctcttcttggatatctggaggaactggcaaaaggat ggtgacatgaaaatcctgcagagc cagattatctctttctacctcagactctttgaagtcttgaaagacaatcaggccatcagc aacaacataagcgtcattgaatcacacc tgattactaccttcttcagcaacagcaaggcgaaaaaggatgcattcatgagtattgcca agtttgaggtcaacaacccacaggtc cagcgccaagcattcaatgagctcatccgagtggtccaccagctgttgccggaatccagc ctcaggaagcggaaaaggagtcg ctgctgaGaattcacgcgttaagtcgacaatcaacctctggattacaaaatttgtgaaag attgactggtattcttaactatgttgctc cttttacgctatgtggatacgctgctttaatgcctttgtatcatgctattgcttcccgta tggctttcattttctcctccttgtataaatcctg gttgctgtctctttatgaggagttgtggcccgttgtcaggcaacgtggcgtggtgtgcac tgtgtttgctgacgcaacccccactgg ttggggcattgccaccacctgtcagctcctttccgggactttcgctttccccctccctat tgccacggcggaactcatcgccgcctg ccttgcccgctgctggacaggggctcggctgttgggcactgacaattccgtggtgttgtc ggggaaatcatcgtcctttccttggct gctcgcctgtgttgccacctggattctgcgcgggacgtccttctgctacgtcccttcggc cctcaatccagcggaccttccttcccg cggcctgctgccggctctgcggcctcttccgcgtcttcgccttcgccctcagacgagtcg gatctccctttgggccgcctccccg cgtcgactttaagaccaatgacGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCC T

CTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACG

CCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTG

CAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACA

CCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCG

CGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTT

AGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTT

TCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTT

TACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGG

GCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTT

TAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCT

ATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATG

AGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACA

ATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCA

GCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCC

CGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCA

GAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATA

CGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGG

TGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAAT

ACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAAT

AATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATT

CCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTG

AAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAAC TGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTT

CCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTAT

TGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGAC

TTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAG

TAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAA

CTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACA

AC AT GGGGGATC ATGT AACTCGCCTT GATCGTT GGGAACCGGAGCTGAAT GA

AGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACA

ACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACA

ATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCG

GCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGG

AAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATC

GT AGTT ATCTACAC G AC GGGG AGT C AGGC A AC TAT GG AT G A AC G A A AT AG AC

AGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCA

AGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAG

GATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTG

AGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCT

TGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACC

GCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGA

AGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAG

CCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGC

TCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTA

C C GGGTT GG AC T C A AG AC GAT AGT T AC C GG AT A AGGC GC AGC GGT C GGGC T G

AACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAA

CTGAGAT ACCT AC AGCGT GAGCT AT GAGAAAGCGCC ACGCTTCCCGAAGGGA

GAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCA

CGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTT

CGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAG

CCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCT

GGCCTTTTGCTCACATGT pGW127b-pAAV EFS-LTR-hIFNg-P2A-hIL23-T2A-h41BBL-WPRE (SEQ ID NO: 49)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGAAATATACAAGTTATATCTTGGCTTTTCAGCTCTGCATCGTTTTGGGTTCT

CTTGGCTGTTACTGCCAGGACCCATATGTAAAAGAAGCAGAAAACCTTAAGA

AATATTTTAATGCAGGTCATTCAGATGTAGCGGATAATGGAACTCTTTTCTTA

GGCATTTTGAAGAATTGGAAAGAGGAGAGTGACAGAAAAATAATGCAGAGC CAAATTGTCTCCTTTTACTTCAAACTTTTTAAAAACTTTAAAGATGACCAGAG

C ATCC AAAAGAGT GT GGAGACC ATC AAGGAAGAC AT GAAT GT C AAGTTTTT C

AAT AGC A AC AAAAAGAAACGAGATGACTTCGAAAAGCTGACT AATT ATTCGG

T AACTGACTT GAAT GTCC AACGC AAAGC AAT AC AT GAACTC ATCC AAGT GAT

GGCTGAACTGTCGCCAGCAGCTAAAACAGGGAAGCGAAAAAGGAGTCAGAT

GCTGTTTCGAGGTCGAAGAGCATCCCAGggcggaGGATCCGGCGCAACAAACTT

CTCTCTGCTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGATGTGT

CACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCCCCTC

GT GGCC AT AT GGGAACTGAAGAAAGAT GTTT AT GTCGT AGAATTGGATT GGT

ATCCGGATGCCCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGA

AG AT GGT ATC ACCTGGACCTTGGACC AGAGC AGT GAGGTCTT AGGCTCTGGC

AAAACCCTGACCATCCAAGTCAAAGAGTTTGGAGATGCTGGCCAGTACACCT

GTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACAAAAA

GGAAGATGGA ATTT GGTCC ACTGAT ATTTT AAAGGACC AGAAAGA ACCC AAA

AATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTG

CTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGC

AGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTCTCTGC

AGAGAGAGTC AGAGGGG AC A AC A AGGAGT AT GAGT ACTC AGT GGAGT GC C A

GGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGGTCATG

GTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCTTCAT

CAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTA

AAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTA

CTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGC

A AGAG AGA A A AGA A AGAT AG AGT C TT C ACGG AC A AGAC CTC AGC C AC GGT C

ATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCTACTATA

GCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTGGCactAGTGGAGAG

GGC AGAGGAAGTCTGCT A AC AT GCGGT GACGTCGAGGAGAATCCTGGCCC AA

TGGAATACGCCTCTGACGCTTCACTGGACCCCGAAGCCCCGTGGCCTCCCGCG

CCCCGCGCTCGCGCCTGCCGCGTACTGCCTTGGGCCCTGGTCGCGGGGCTGCT

GCTGCTGCTGCTGCTCGCTGCCGCCTGCGCCGTCTTCCTCGCCTGCCCCTGGG

CCGTGTCCGGGGCTCGCGCCTCGCCCGGCTCCGCGGCCAGCCCGAGACTCCG

CGAGGGTCCCGAGCTTTCGCCCGACGATCCCGCCGGCCTCTTGGACCTGCGGC

AGGGCATGTTTGCGCAGCTGGTGGCCCAAAATGTTCTGCTGATCGATGGGCCC

CTGAGCTGGTACAGTGACCCAGGCCTGGCAGGCGTGTCCCTGACGGGGGGCC

T GAGCT AC AAAGAGGAC ACGAAGGAGCTGGT GGT GGCC AAGGCTGGAGTCT

ACTATGTCTTCTTTCAACTAGAGCTGCGGCGCGTGGTGGCCGGCGAGGGCTCA

GGCTCCGTTTCACTTGCGCTGCACCTGCAGCCACTGCGCTCTGCTGCTGGGGC

CGCCGCCCTGGCTTTGACCGTGGACCTGCCACCCGCCTCCTCCGAGGCTCGGA

ACTCGGCCTTCGGTTTCCAGGGCCGCTTGCTGCACCTGAGTGCCGGCCAGCGC

CTGGGCGTCCATCTTCACACTGAGGCCAGGGCACGCCATGCCTGGCAGCTTA

CCCAGGGCGCCACAGTCTTGGGACTCTTCCGGGTGACCCCCGAAATCCCAGC

CGGACTCCCTT C ACCGAGGTCGGAAT AAGaattcacgcgttaagtcgacaatcaacctctggattaca aaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgtggat acgctgctttaatgcctttgtatcatgctattg cttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctctttatg aggagttgtggcccgttgtcaggcaacgtgg cgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccacctg tcagctcctttccgggactttcgctttc cccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggacaggg gctcggctgttgggcactgacaatt ccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgccacct ggattctgcgcgggacgtccttctgcta cgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctctgcg gcctcttccgcgtcttcgccttcgcc ctcagacgagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaatgac GGCCGCAGGAACCCC

TAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCC GGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGA

GCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTT

ACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGC

GCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTG

ACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCC

TTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCC

TTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATT

TGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCT

TTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAAC

AACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGAT

TTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATT

TTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGC

TCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACG

CGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACC

GTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGC

GAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAA

TAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACC

CCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAA

T A ACCC T GAT A A AT GCTT C A AT A AT ATT GA A A A AGGA AGAGT AT GAGT ATTC

AACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTT

T GCTC ACCC AGAAACGCTGGTGAA AGT A AAAGAT GCTGAAGAT C AGTTGGGT

GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGA

GTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTA

TGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCC

GCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA

GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACC

ATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA

AGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGAT

CGTTGGGAACCGGAGCTGAAT GAAGCC AT ACC AAACGACGAGCGT GAC ACC A

CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACT

ACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAA

GTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGA

T A A AT C T GG AGC C GGT G AGC GT GG A AGC C GC GGT AT C ATT GC AGC AC T GGGG

CCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG

CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGAT

TAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATT

TAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAAT

CTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCC

CGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCT

GCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGA

TCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAG

ATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA

CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTG

CTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTA

CCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCA

GCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATG

AGAAAGCGCC ACGCTTCCCGAAGGGAGAAAGGCGGAC AGGT ATCCGGT AAG

CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGC

CTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATT TTTGT GAT GCTCGT C AGGGGGGCGGAGCCT AT GGAAAAACGCC AGC AACGCG GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW128b-pAAV EFS-LTR-hIFNg-P2A-hIL23-T2A-h41BBL-E2A-hLight-WPRE (SEQ ID NO: 50)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGAAATATACAAGTTATATCTTGGCTTTTCAGCTCTGCATCGTTTTGGGTTCT

CTTGGCTGTTACTGCCAGGACCCATATGTAAAAGAAGCAGAAAACCTTAAGA

AATATTTTAATGCAGGTCATTCAGATGTAGCGGATAATGGAACTCTTTTCTTA

GGCATTTTGAAGAATTGGAAAGAGGAGAGTGACAGAAAAATAATGCAGAGC

CAAATTGTCTCCTTTTACTTCAAACTTTTTAAAAACTTTAAAGATGACCAGAG

C ATCC AAAAGAGT GT GGAGACC ATC AAGGAAGAC AT GAAT GTC AAGTTTTT C

AAT AGC A AC AAAAAGAAACGAGATGACTTCGAAAAGCTGACT AATT ATTCGG

T AACTGACTT GAAT GTCC A AC GCA A AGC A AT AC AT GAACTC ATCC AAGT GAT

GGCTGAACTGTCGCCAGCAGCTAAAACAGGGAAGCGAAAAAGGAGTCAGAT

GCTGTTTCGAGGTCGAAGAGCATCCCAGggcggaGGATCCGGCGCAACAAACTT

CTCTCTGCTGAAACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGATGTGT

CACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTCCCCTC

GT GGCC AT AT GGGAACTGAAGAAAGAT GTTT AT GTCGT AGAATTGGATT GGT

ATCCGGATGCCCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTGAAGA

AG AT GGT ATC ACCTGGACCTTGGACC AGAGC AGT GAGGTCTT AGGCTCTGGC

AAAACCCTGACCATCCAAGTCAAAGAGTTTGGAGATGCTGGCCAGTACACCT

GTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACAAAAA

GGAAGATGGA ATTT GGTCC ACTGAT ATTTT AAAGGACC AGAAAGA ACCC AAA

AATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCACCTG

CTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGCAGC

AGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTCTCTGC

AGAGAGAGTC AGAGGGG AC A AC A AGGAGT AT GAGT ACTC AGT GGAGT GC C A

GGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGGTCATG

GTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTCTTCAT

CAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGCCATTA

AAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTGGAGTA

CTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGCAAGAGC

A AGAG AGA A A AGA A AGAT AG AGT C TT C ACGG AC A AGAC CTC AGC C AC GGT C

ATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCTACTATA

GCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTGGCAGTGGAGAGGG

CAGAGGAAGTCTGCTAACATGCGGTGACGTCGAGGAGAATCCTGGCCCAATG

GAATACGCCTCTGACGCTTCACTGGACCCCGAAGCCCCGTGGCCTCCCGCGCC CCGCGCTCGCGCCTGCCGCGTACTGCCTTGGGCCCTGGTCGCGGGGCTGCTGC

TGCTGCTGCTGCTCGCTGCCGCCTGCGCCGTCTTCCTCGCCTGCCCCTGGGCC

GTGTCCGGGGCTCGCGCCTCGCCCGGCTCCGCGGCCAGCCCGAGACTCCGCG

AGGGTCCCGAGCTTTCGCCCGACGATCCCGCCGGCCTCTTGGACCTGCGGCA

GGGCATGTTTGCGCAGCTGGTGGCCCAAAATGTTCTGCTGATCGATGGGCCCC

TGAGCTGGTACAGTGACCCAGGCCTGGCAGGCGTGTCCCTGACGGGGGGCCT

GAGCTACAAAGAGGACACGAAGGAGCTGGTGGTGGCCAAGGCTGGAGTCTA

CTATGTCTTCTTTCAACTAGAGCTGCGGCGCGTGGTGGCCGGCGAGGGCTCAG

GCTCCGTTTCACTTGCGCTGCACCTGCAGCCACTGCGCTCTGCTGCTGGGGCC

GCCGCCCTGGCTTTGACCGTGGACCTGCCACCCGCCTCCTCCGAGGCTCGGAA

CTCGGCCTTCGGTTTCCAGGGCCGCTTGCTGCACCTGAGTGCCGGCCAGCGCC

TGGGCGTCCATCTTCACACTGAGGCCAGGGCACGCCATGCCTGGCAGCTTAC

CCAGGGCGCCACAGTCTTGGGACTCTTCCGGGTGACCCCCGAAATCCCAGCC

GGACTCCCTTCACCGAGGTCGGAAGGTgctagcggtacccagtgcaccaactacgcc ctgctgaagct ggccggcgatgtggagagcaaccccgggccc AT GGAGGAGAGT GTCGT ACGGCCCTC AGT GTT

TGTGGTGGATGGACAGACCGACATCCCATTCACGAGGCTGGGACGAAGCCAC

CGGAGACAGTCGTGCAGTGTGGCCCGGGTGGGTCTGGGTCTCTTGCTGTTGCT

GATGGGGGCCGGGCTGGCCGTCCAAGGCTGGTTCCTCCTGCAGCTGCACTGG

CGTCTAGGAGAGATGGTCACCCGCCTGCCTGACGGACCTGCAGGCTCCTGGG

AGCAGCTGATACAAGAGCGAAGGTCTCACGAGGTCAACCCAGCAGCGCATCT

CACAGGGGCCAACTCCAGCTTGACCGGCAGCGGGGGGCCGCTGTTATGGGAG

ACTCAGCTGGGCCTGGCCTTCCTGAGGGGCCTCAGCTACCACGATGGGGCCC

TTGTGGTCACCAAAGCTGGCTACTACTACATCTACTCCAAGGTGCAGCTGGGC

GGTGTGGGCTGCCCGCTGGGCCTGGCCAGCACCATCACCCACGGCCTCTACA

AGCGCACACCCCGCTACCCCGAGGAGCTGGAGCTGTTGGTCAGCCAGCAGTC

ACCCTGCGGACGGGCCACCAGCAGCTCCCGGGTCTGGTGGGACAGCAGCTTC

C T GGGT GGT GT GGT AC AC C T GG AGGC T GGGG AG A AGGT GGT C GT C C GT GT GC

TGGATGAACGCCTGGTTCGACTGCGTGATGGTACCCGGTCTTACTTCGGGGCT

TTC AT GGT GGGTGGAGGCGGTTGCGGAA AGCGT AAGT GAGaattcacgcgttaagtcgac aatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgct ccttttacgctatgtggatacgctgctttaat gcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttgtataaatc ctggttgctgtctctttatgaggagttgtggcc cgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttg gggcattgccaccacctgtcagctc ctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccgcctgc cttgcccgctgctggacaggggctcg gctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttccttggct gctcgcctgtgttgccacctggattctg cgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcccgc ggcctgctgccggctctgcggcctct tccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctccccgcg tcgactttaagaccaatgacGGC

CGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCG

CTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGG

CGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCG

GTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCA

ACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTT

ACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGC

TTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAA

TCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCA

AAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGAC

GGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTT

CCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAG

GGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAAAA

TTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCT

CAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCA ACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAG

ACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCAT

CACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT

TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAA

ATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC

CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA

GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT

TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTG

AAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGG

TAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT

TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAG

CAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACC

AGT C AC AGAAAAGC ATCTT ACGGAT GGC ATGAC AGT AAGAGAATT AT GC AGT

GCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGAT

CGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA

ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG

AGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATT

AACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGG

AGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTG

GTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTG

CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGAC

GGGGAGTC AGGC A ACT AT GGAT GA ACGA A AT AGAC AGATCGC T GAG AT AGG

TGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATAC

TTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATC

CTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGA

GCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCT

GCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTT

TGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAG

CAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACC

ACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTA

CCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAG

ACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGC

ACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGC

GTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGT

ATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAG

GGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT

GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACG

CCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACA

TGT pGW145-pAAV EFS-LTR-hIFNg-WPRE-sPA (SEQ ID NO: 51)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGAAATATACAAGTTATATCTTGGCTTTTCAGCTCTGCATCGTTTTGGGTTCT

CTTGGCTGTTACTGCCAGGACCCATATGTAAAAGAAGCAGAAAACCTTAAGA

AATATTTTAATGCAGGTCATTCAGATGTAGCGGATAATGGAACTCTTTTCTTA

GGCATTTTGAAGAATTGGAAAGAGGAGAGTGACAGAAAAATAATGCAGAGC

CAAATTGTCTCCTTTTACTTCAAACTTTTTAAAAACTTTAAAGATGACCAGAG

C ATCC AAAAGAGT GT GGAGACC ATC AAGGAAGAC AT GAAT GT C AAGTTTTT C

AAT AGC A AC AAAAAGAAACGAGATGACTTCGAAAAGCTGACT AATT ATTCGG

T AACTGACTT GAAT GTCC AACGC AAAGC AAT AC AT GAACTC ATCC AAGT GAT

GGCTGAACTGTCGCCAGCAGCTAAAACAGGGAAGCGAAAAAGGAGTCAGAT

GCTGTTTCGAGGTCGAAGAGC ATCCC AGT GAGaattcacgcgttaagtcgacaatcaacctctgga ttacaaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatg tggatacgctgctttaatgcctttgtatcatgc tattgcttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctct ttatgaggagttgtggcccgttgtcaggcaac gtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccacca cctgtcagctcctttccgggactttcg ctttccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctgga caggggctcggctgttgggcactga caattccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgc cacctggattctgcgcgggacgtccttct gctacgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctc tgcggcctcttccgcgtcttcgccttc gccctcagacgagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaat gacGGCCGaataaaagatcttta ttttcattagatctgtgtgttggttttttgtgtgcggccgCAGGAACCCCTAGTGATGGA GTTGGCCACT

CCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCC

GACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTG

CCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTT

CACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTA

AGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCG

CCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCG

GCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGT

GCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAG

TGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGT

TCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCG

GGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAA

AATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTT

TACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAA

GCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCT

GCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGT

GTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGT

GATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGT

CAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTC

TAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCT

TCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCC

TTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGC

T GGT GA A AGT A A A AGAT GC T GA AG ATC AGTT GGGT GC AC GAGT GGGTT AC AT

CGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAA

CGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATC

CCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAG

AATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCA

T GAC AGT A AG AG A ATT AT GC AGT GC T GC CAT A ACC AT GAGT GAT A AC AC T GC

GGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTT TTGC AC AAC AT GGGGGAT CAT GT AACTCGCCTT GATCGTT GGGAACCGGAGC

TGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAAT

GGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCC

GGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCT

GCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTG

AGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTC

CCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGA

AATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGT

CAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAAT

TTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCT

TAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAG

GATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAA

AACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTT

TTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCT

AGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACAT

ACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCG

TGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT

CGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTA

CACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCC

GA AGGGAGA A AGGC GGAC AGGT ATCC GGT A AGC GGC AGGGT C GGA AC AGGA

GAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTG

TCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGG

GGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGG

CCTTTTGCTGGCCTTTTGCTCACATGT pGW146-pAAV EFS-LTR-hIL23-WPRE-sPA (SEQ ID NO: 52)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGTGTCACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCT

CCCCTCGTGGCCATATGGGAACTGAAGAAAGATGTTTATGTCGTAGAATTGG

ATTGGTATCCGGATGCCCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCT

GAAGAAGATGGTATCACCTGGACCTTGGACCAGAGCAGTGAGGTCTTAGGCT

CTGGCAAAACCCTGACCATCCAAGTCAAAGAGTTTGGAGATGCTGGCCAGTA

CACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACA

AAAAGGAAGAT GGAATTTGGTCC ACTGAT ATTTT AAAGGACC AGAAAGA ACC

CAAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTC

ACCTGCTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAG

CAGCAGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTC

TCTGC AGAGAGAGT C AG AGGGGAC A AC A AGG AGT AT GAGT ACTC AGT GGAG TGCCAGGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGG

T CAT GGTGGAT GCCGTT C AC AAGCTC AAGT AT GAAAACT AC ACC AGC AGCTT

CTTCATCAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAG

CCATTAAAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCT

GGAGTACTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGC

A AGAGC A AGAGAGA A A AGA A AGAT AGAGT C TTC ACGGAC A AGAC CTC AGC C

ACGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCT

ACTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGTTGAGaattcacg cgttaagtcgacaatcaacctctggattacaaaatttgtgaaagattgactggtattctt aactatgttgctccttttacgctatgtggat acgctgctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctcct ccttgtataaatcctggttgctgtctctttatgag gagttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacc cccactggttggggcattgccacca cctgtcagctcctttccgggactttcgctttccccctccctattgccacggcggaactca tcgccgcctgccttgcccgctgctgga caggggctcggctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcct ttccttggctgctcgcctgtgttgcca cctggattctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcggacc ttccttcccgcggcctgctgccggct ctgcggcctcttccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggcc gcctccccgcgtcgactttaagacca atgacGGCCGaataaaagatctttattttcattagatctgtgtgttggttttttgtgtgc ggccgCAGGAACCCCTAG

TGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGG

CGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCG

AGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACG

CATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCC

TGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCG

CTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTC

TCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTA

GGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGG

TGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGA

CGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACA

CTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCG

GTCT ATT GGTT AAAAAAT GAGCTGATTT AAC AAAAATTT AACGCGAATTTT AA

CAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTG

ATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCC

CTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCT

CCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAG

ACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAA

TGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCT

ATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA

CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAAC

ATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGC

TCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCA

CGAGTGGGTT AC ATCGAACTGGATCTC AAC AGCGGT AAGATCCTT GAGAGTT

TTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGT

GGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCA

TACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCAT

CTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGA

GTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGA

GCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTT

GGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGA

TGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTT

ACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTG

CAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAA

TCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAG ATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAAC

TATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAG

CATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAA

ACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCAT

GACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAG

AAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGC

TTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAG

AGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCA

AATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGT

AGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCA

GTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGA

TAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTG

GAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAA

GCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCA

GGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGT

ATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGT

GATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTT

TTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW147-pAAV EFS-LTR-h41BBL-WPRE-sPA (SEQ ID NO: 53)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGGAATACGCCTCTGACGCTTCACTGGACCCCGAAGCCCCGTGGCCTCCCGC

GCCCCGCGCTCGCGCCTGCCGCGTACTGCCTTGGGCCCTGGTCGCGGGGCTGC

TGCTGCTGCTGCTGCTCGCTGCCGCCTGCGCCGTCTTCCTCGCCTGCCCCTGG

GCCGTGTCCGGGGCTCGCGCCTCGCCCGGCTCCGCGGCCAGCCCGAGACTCC

GCGAGGGTCCCGAGCTTTCGCCCGACGATCCCGCCGGCCTCTTGGACCTGCG

GCAGGGCATGTTTGCGCAGCTGGTGGCCCAAAATGTTCTGCTGATCGATGGG

CCCCTGAGCTGGTACAGTGACCCAGGCCTGGCAGGCGTGTCCCTGACGGGGG

GCCTGAGCTACAAAGAGGACACGAAGGAGCTGGTGGTGGCCAAGGCTGGAG

TCTACTATGTCTTCTTTCAACTAGAGCTGCGGCGCGTGGTGGCCGGCGAGGGC

TCAGGCTCCGTTTCACTTGCGCTGCACCTGCAGCCACTGCGCTCTGCTGCTGG

GGCCGCCGCCCTGGCTTTGACCGTGGACCTGCCACCCGCCTCCTCCGAGGCTC

GGAACTCGGCCTTCGGTTTCCAGGGCCGCTTGCTGCACCTGAGTGCCGGCCAG

CGCCTGGGCGTCCATCTTCACACTGAGGCCAGGGCACGCCATGCCTGGCAGC

TTACCCAGGGCGCCACAGTCTTGGGACTCTTCCGGGTGACCCCCGAAATCCCA

GCCGGACTCCCTTCACCGAGGTCGGAATGAGaattcacgcgttaagtcgacaatcaa cctctggatt acaaaatttgtgaaagattgactggtattcttaactatgttgctccttttacgctatgtg gatacgctgctttaatgcctttgtatcatgcta ttgcttcccgtatggctttcattttctcctccttgtataaatcctggttgctgtctcttt atgaggagttgtggcccgttgtcaggcaacgt ggcgtggtgtgcactgtgtttgctgacgcaacccccactggttggggcattgccaccacc tgtcagctcctttccgggactttcgct ttccccctccctattgccacggcggaactcatcgccgcctgccttgcccgctgctggaca ggggctcggctgttgggcactgaca attccgtggtgttgtcggggaaatcatcgtcctttccttggctgctcgcctgtgttgcca cctggattctgcgcgggacgtccttctg ctacgtcccttcggccctcaatccagcggaccttccttcccgcggcctgctgccggctct gcggcctcttccgcgtcttcgccttcg ccctcagacgagtcggatctccctttgggccgcctccccgcgtcgactttaagaccaatg acGGCCGaataaaagatctttatt ttcattagatctgtgtgttggttttttgtgtgcggccgC AGGA AC CCC T AGT GAT GGAGTTGGCC AC T C

CCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCG

ACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGC

CTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTC

ACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAA

GCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGC

CTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGG

CTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG

CTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGT

GGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTT

CTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGG

GCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAA

ATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTT

ACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAG

CCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTG

CTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTG

TCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTG

ATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTC

AGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCT

AAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTT

C AAT AAT ATT GAAAAAGGA AGAGT ATGAGT ATT C A AC ATTTCCGT GTCGCCCT

TATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCT

GGT GA A AGT A A A AG AT GCTGA AGAT C AGTT GGGT GC ACGAGT GGGTT AC AT C

GAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAAC

GTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCC

CGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGA

ATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT

GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCG

GCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTT

GCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTG

AATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGG

CAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGG

C AAC AATT AAT AGACTGGAT GGAGGCGGAT AA AGTTGC AGGACC ACTTCTGC

GCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAG

CGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCC

GTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAA

TAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCA

GACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTT

AAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTA

ACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGA

TCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAA

CCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTT

TCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAG

TGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATAC

CTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTG TCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCG

GGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACA

CCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGA

AGGGAGA A AGGC GGAC AGGT ATCCGGT A AGC GGC AGGGTCGGA AC AGGAGA

GCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTC

GGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGG

GCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCC

TTTTGCTGGCCTTTTGCTCACATGT pGW149-pAAV EFS-LTR-hLIGHT(Remove-EQLI)-RSR-WPRE-sPA (SEQ ID NO: 54)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c

ATGGAGGAGAGTGTCGTACGGCCCTCAGTGTTTGTGGTGGATGGACAGACCG

ACATCCCATTCACGAGGCTGGGACGAAGCCACCGGAGACAGTCGTGCAGTGT

GGCCCGGGTGGGTCTGGGTCTCTTGCTGTTGCTGATGGGGGCCGGGCTGGCCG

TCCAAGGCTGGTTCCTCCTGCAGCTGCACTGGCGTCTAGGAGAGATGGTCACC

CGCCTGCCTGACGGACCTGCAGGCTCCTGGCAAGAGCGAAGGTCTCACGAGG

TCAACCCAGCAGCGCATCTCACAGGGGCCAACTCCAGCTTGACCGGCAGCGG

GGGGCCGCTGTTATGGGAGACTCAGCTGGGCCTGGCCTTCCTGAGGGGCCTC

AGCTACCACGATGGGGCCCTTGTGGTCACCAAAGCTGGCTACTACTACATCTA

CTCCAAGGTGCAGCTGGGCGGTGTGGGCTGCCCGCTGGGCCTGGCCAGCACC

ATCACCCACGGCCTCTACAAGCGCACACCCCGCTACCCCGAGGAGCTGGAGC

TGTTGGTCAGCCAGCAGTCACCCTGCGGACGGGCCACCAGCAGCTCCCGGGT

CTGGT GGGAC AGC AGCTTCCTGGGTGGT GT GGT AC ACCTGGAGGCTGGGGAG

AAGGTGGTCGTCCGTGTGCTGGATGAACGCCTGGTTCGACTGCGTGATGGTAC

CCGGTCTTACTTCGGGGCTTTCATGGTGggtggaggcggttgcggaaagcgtaagTG AGaattca cgcgttaagtcgacaatcaacctctggattacaaaatttgtgaaagattgactggtattc ttaactatgttgctccttttacgctatgtgg atacgctgctttaatgcctttgtatcatgctattgcttcccgtatggctttcattttctc ctccttgtataaatcctggttgctgtctctttatg aggagttgtggcccgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaa cccccactggttggggcattgccac cacctgtcagctcctttccgggactttcgctttccccctccctattgccacggcggaact catcgccgcctgccttgcccgctgctg gacaggggctcggctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtc ctttccttggctgctcgcctgtgttgc cacctggattctgcgcgggacgtccttctgctacgtcccttcggccctcaatccagcgga ccttccttcccgcggcctgctgccgg ctctgcggcctcttccgcgtcttcgccttcgccctcagacgagtcggatctccctttggg ccgcctccccgcgtcgactttaagacc aatgacGGCCGaataaaagatctttattttcattagatctgtgtgttggttttttgtgtg cggccgCAGGAACCCCTAG

TGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGG

CGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCG

AGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACG

CATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCC TGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCG

CTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTC

TCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTA

GGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGG

TGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGA

CGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACA

CTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCG

GTCT ATT GGTT AAAAAAT GAGCTGATTT AAC AAAAATTT AACGCGAATTTT AA

CAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTG

ATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCC

CTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCT

CCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAG

ACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAA

TGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCT

ATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA

CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAAC

ATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGC

TCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCA

CGAGTGGGTT AC ATCGAACTGGATCTC AAC AGCGGT AAGATCCTT GAGAGTT

TTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGT

GGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCA

TACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCAT

CTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGA

GTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGA

GCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTT

GGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGA

TGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTT

ACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTG

CAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAA

TCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCAGCACTGGGGCCAG

ATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAAC

TATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAG

CATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAA

ACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCAT

GACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAG

AAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGC

TTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAG

AGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCA

AATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGT

AGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCA

GTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGA

TAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTG

GAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAA

GCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCA

GGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGT

ATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGT

GATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTT

TTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW036-EFS-dSaCas9-VP64 (SEQ ID NO: 55) CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atggccccaaagaagaagcggaaggtcggtatccacggagtcccagcagccaagcggaac tacatcctgggcctggccatc ggcatcaccagcgtgggctacggcatcatcgactacgagacacgggacgtgatcgatgcc ggcgtgcggctgttcaaagagg ccaacgtggaaaacaacgagggcaggcggagcaagagaggcgccagaaggctgaagcggc ggaggcggcatagaatcc agagagtgaagaagctgctgttcgactacaacctgctgaccgaccacagcgagctgagcg gcatcaacccctacgaggccag agtgaagggcctgagccagaagctgagcgaggaagagttctctgccgccctgctgcacct ggccaagagaagaggcgtgca caacgtgaacgaggtggaagaggacaccggcaacgagctgtccaccaaagagcagatcag ccggaacagcaaggccctgg aagagaaatacgtggccgaactgcagctggaacggctgaagaaagacggcgaagtgcggg gcagcatcaacagattcaaga ccagcgactacgtgaaagaagccaaacagctgctgaaggtgcagaaggcctaccaccagc tggaccagagcttcatcgacac ctacatcgacctgctggaaacccggcggacctactatgagggacctggcgagggcagccc cttcggctggaaggacatcaaa gaatggtacgagatgctgatgggccactgcacctacttccccgaggaactgcggagcgtg aagtacgcctacaacgccgacct gtacaacgccctgaacgacctgaacaatctcgtgatcaccagggacgagaacgagaagct ggaatattacgagaagttccaga tcatcgagaacgtgttcaagcagaagaagaagcccaccctgaagcagatcgccaaagaaa tcctcgtgaacgaagaggatatt aagggctacagagtgaccagcaccggcaagcccgagttcaccaacctgaaggtgtaccac gacatcaaggacattaccgccc ggaaagagattattgagaacgccgagctgctggatcagattgccaagatcctgaccatct accagagcagcgaggacatccag gaagaactgaccaatctgaactccgagctgacccaggaagagatcgagcagatctctaat ctgaagggctataccggcaccca caacctgagcctgaaggccatcaacctgatcctggacgagctgtggcacaccaacgacaa ccagatcgctatcttcaaccggct gaagctggtgcccaagaaggtggacctgtcccagcagaaagagatccccaccaccctggt ggacgacttcatcctgagcccc gtcgtgaagagaagcttcatccagagcatcaaagtgatcaacgccatcatcaagaagtac ggcctgcccaacgacatcattatcg agctggcccgcgagaagaactccaaggacgcccagaaaatgatcaacgagatgcagaagc ggaaccggcagaccaacgag cggatcgaggaaatcatccggaccaccggcaaagagaacgccaagtacctgatcgagaag atcaagctgcacgacatgcag gaaggcaagtgcctgtacagcctggaagccatccctctggaagatctgctgaacaacccc ttcaactatgaggtggaccacatc atccccagaagcgtgtccttcgacaacagcttcaacaacaaggtgctcgtgaagcaggaa gaagccagcaagaagggcaacc ggaccccattccagtacctgagcagcagcgacagcaagatcagctacgaaaccttcaaga agcacatcctgaatctggccaag ggcaagggcagaatcagcaagaccaagaaagagtatctgctggaagaacgggacatcaac aggttctccgtgcagaaagact tcatcaaccggaacctggtggataccagatacgccaccagaggcctgatgaacctgctgc ggagctacttcagagtgaacaac ctggacgtgaaagtgaagtccatcaatggcggcttcaccagctttctgcggcggaagtgg aagtttaagaaagagcggaacaa ggggtacaagcaccacgccgaggacgccctgatcattgccaacgccgatttcatcttcaa agagtggaagaaactggacaagg ccaaaaaagtgatggaaaaccagatgttcgaggaaaagcaggccgagagcatgcccgaga tcgaaaccgagcaggagtaca aagagatcttcatcaccccccaccagatcaagcacattaaggacttcaaggactacaagt acagccaccgggtggacaagaag cctaatagagagctgattaacgacaccctgtactccacccggaaggacgacaagggcaac accctgatcgtgaacaatctgaa cggcctgtacgacaaggacaatgacaagctgaaaaagctgatcaacaagagccccgaaaa gctgctgatgtaccaccacgac ccccagacctaccagaaactgaagctgattatggaacagtacggcgacgagaagaatccc ctgtacaagtactacgaggaaac cgggaactacctgaccaagtactccaaaaaggacaacggccccgtgatcaagaagattaa gtattacggcaacaaactgaacg cccatctggacatcaccgacgactaccccaacagcagaaacaaggtcgtgaagctgtccc tgaagccctacagattcgacgtgt acctggacaatggcgtgtacaagttcgtgaccgtgaagaatctggatgtgatcaaaaaag aaaactactacgaagtgaatagcaa gtgctatgaggaagctaagaagctgaagaagatcagcaaccaggccgagtttatcgcctc cttctacaacaacgatctgatcaag atcaacggcgagctgtatagagtgatcggcgtgaacaacgacctgctgaaccggatcgaa gtgaacatgatcgacatcacctac cgcgagtacctggaaaacatgaacgacaagaggccccccaggatcattaagacaatcgcc tccaagacccagagcattaaga agtacagcacagacattctgggcaacctgtatgaagtgaaatctaagaagcaccctcaga tcatcaaaaagggcaaaaggccg gcggccacgaaaaaggccggccaggcaaaaaagaaaaagggatccGGACGGGCTGACGCA TTGGACG

ATTTTGATCTGGATATGCTGGGAAGTGACGCCCTCGATGATTTTGACCTTGAC

ATGCTTGGTTCGGATGCCCTTGATGACTTTGACCTCGACATGCTCGGCAGTGA

CGCCCTTGATGATTTCGACCTGGACATGCTGATTTAAGAATTCAATAAAAGAT

CTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCGGCCGCAGGAACCC

CTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGC

CGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTG

AGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCC

TTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACG

CGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGT

GACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTC

CTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCC

CTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGAT

TTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCC

TTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAA

CAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGA

TTTCGGTCT ATTGGTT AAAAAAT GAGCTGATTT AAC AA AAATTT AACGCGAAT

TTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTG

CTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGAC

GCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGAC

CGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCG

CGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATA

ATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAAC

CCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACA

ATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATT

CAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTT

TTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGG

TGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAG

AGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCT

ATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGC

CGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAA

GCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACC

ATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA

AGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGAT

CGTTGGGAACCGGAGCTGAAT GAAGCC AT ACC AAACGACGAGCGT GAC ACC A

CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACT

ACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAA

GTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGA

T A A AT C T GG AGC C GGT G AGC GT GG A AGC C GC GGT AT C ATT GC AGC AC T GGGG

CCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG

CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGAT

TAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATT

TAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAAT

CTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCC

CGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCT

GCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGA

TCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAG

ATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTG

CTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTA

CCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCA

GCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATG

AGAAAGCGCC ACGCTTCCCGAAGGGAGAAAGGCGGAC AGGT ATCCGGT AAG

CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGC

CTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATT

TTTGT GAT GCTCGT C AGGGGGGCGGAGCCT AT GGAAAAACGCC AGC AACGCG

GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGT pGW011b_pAAV-EFS-dCAS9-spA (SEQ ID NO: 56) cctgcaggcagctgcgcgctcgctcgctcactgaggccgcccgggcaaagcccgggcgtc gggcgacctttggtcgcccgg cctcagtgagcgagcgagcgcgcagagagggagtggccaactccatcactaggggttcct gcggccgcacgcgttctaggtc ttgaaaggagtgggaattggctccggtgcccgtcagtgggcagagcgcacatcgcccaca gtccccgagaagttggggggag gggtcggcaattgatccggtgcctagagaaggtggcgcggggtaaactgggaaagtgatg tcgtgtactggctccgcctttttcc cgagggtgggggagaaccgtatataagtgcagtagtcgccgtgaacgttctttttcgcaa cgggtttgccgccagaacacaggT

GTCGTGACGCGcGTACGtaatacgactcactatagggccgccaccATGAAAAGGCCG GCGGCC

ACGAAAAAGGCCGGCCAGGCAAAAAAGAAAAAGGACAAGAAGTACAGCATC

GGCCTGGCCATCGGCACCAACTCTGTGGGCTGGGCCGTGATCACCGACGAGT

ACAAGGTGCCCAGCAAGAAATTCAAGGTGCTGGGCAACACCGACCGGCACA

GCATCAAGAAGAACCTGATCGGAGCCCTGCTGTTCGACAGCGGCGAAACAGC

CGAGGCCACCCGGCTGAAGAGAACCGCCAGAAGAAGATACACCAGACGGAA

GAACCGGATCTGCT ATCTGC A AGAGATCTTC AGC AACGAGAT GGCC AAGGTG

GACGAC AGCTTCTTCC AC AGACTGGAAGAGTCCTTCCTGGTGGAAGAGGAT A

AGAAGCACGAGCGGCACCCCATCTTCGGCAACATCGTGGACGAGGTGGCCTA

CCACGAGAAGTACCCCACCATCTACCACCTGAGAAAGAAACTGGTGGACAGC

ACCGACAAGGCCGACCTGCGGCTGATCTATCTGGCCCTGGCCCACATGATCA

AGTTCCGGGGCCACTTCCTGATCGAGGGCGACCTGAACCCCGACAACAGCGA

CGTGGACAAGCTGTTCATCCAGCTGGTGCAGACCTACAACCAGCTGTTCGAG

GAAAACCCCATCAACGCCAGCGGCGTGGACGCCAAGGCCATCCTGTCTGCCA

GACTGAGCAAGAGCAGACGGCTGGAAAATCTGATCGCCCAGCTGCCCGGCGA

GAAGAAGAATGGCCTGTTCGGCAACCTGATTGCCCTGAGCCTGGGCCTGACC

CCCAACTTCAAGAGCAACTTCGACCTGGCCGAGGATGCCAAACTGCAGCTGA

GCAAGGACACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGCGA

CCAGTACGCCGACCTGTTTCTGGCCGCCAAGAACCTGTCCGACGCCATCCTGC

TGAGCGACATCCTGAGAGTGAACACCGAGATCACCAAGGCCCCCCTGAGCGC

CTCTATGATCAAGAGATACGACGAGCACCACCAGGACCTGACCCTGCTGAAA

GCTCTCGTGCGGCAGCAGCTGCCTGAGAAGTACAAAGAGATTTTCTTCGACC

AGAGCAAGAACGGCTACGCCGGCTACATTGACGGCGGAGCCAGCCAGGAAG

AGTTCTACAAGTTCATCAAGCCCATCCTGGAAAAGATGGACGGCACCGAGGA

ACTGCTCGT GAAGCTGAAC AGAGAGGACCTGCTGCGGA AGC AGCGGACCTT C

GACAACGGCAGCATCCCCCACCAGATCCACCTGGGAGAGCTGCACGCCATTC

T GCGGCGGC AGGAAGATTTTT ACCC ATTCCTGAAGGAC AACCGGGAA AAGAT

CGAGAAGATCCTGACCTTCCGCATCCCCTACTACGTGGGCCCTCTGGCCAGGG

GAAACAGCAGATTCGCCTGGATGACCAGAAAGAGCGAGGAAACCATCACCC

CCTGGAACTTCGAGGAAGTGGTGGACAAGGGCGCTTCCGCCCAGAGCTTCAT

CGAGCGGATGACCAACTTCGATAAGAACCTGCCCAACGAGAAGGTGCTGCCC

AAGCACAGCCTGCTGTACGAGTACTTCACCGTGTATAACGAGCTGACCAAAG

TGAAATACGTGACCGAGGGAATGAGAAAGCCCGCCTTCCTGAGCGGCGAGCA

GAAAAAGGCCATCGTGGACCTGCTGTTCAAGACCAACCGGAAAGTGACCGTG AAGCAGCTGAAAGAGGACTACTTCAAGAAAATCGAGTGCTTCGACTCCGTGG

AAATCTCCGGCGTGGAAGATCGGTTCAACGCCTCCCTGGGCACATACCACGA

TCTGCTGAAAATTATCAAGGACAAGGACTTCCTGGACAATGAGGAAAACGAG

GACATTCTGGAAGATATCGTGCTGACCCTGACACTGTTTGAGGACAGAGAGA

TGATCGAGGAACGGCTGAAAACCTATGCCCACCTGTTCGACGACAAAGTGAT

GAAGCAGCTGAAGCGGCGGAGATACACCGGCTGGGGCAGGCTGAGCCGGAA

GCTGATCAACGGCATCCGGGACAAGCAGTCCGGCAAGACAATCCTGGATTTC

CTGAAGTCCGACGGCTTCGCCAACAGAAACTTCATGCAGCTGATCCACGACG

ACAGCCTGACCTTTAAAGAGGACATCCAGAAAGCCCAGGTGTCCGGCCAGGG

CGATAGCCTGCACGAGCACATTGCCAATCTGGCCGGCAGCCCCGCCATTAAG

A AGGGC ATCCTGC AGAC AGT G A AGGT GGT GGACGAGC T C GT GA A AGT GAT GG

GCCGGCACAAGCCCGAGAACATCGTGATCGAAATGGCCAGAGAGAACCAGA

CCACCCAGAAGGGACAGAAGAACAGCCGCGAGAGAATGAAGCGGATCGAAG

AGGGC ATCAAAGAGCTGGGCAGCCAGATCCTGAAAGAACACCCCGTGG AAA

ACACCCAGCTGCAGAACGAGAAGCTGTACCTGTACTACCTGCAGAATGGGCG

GGATATGTACGTGGACCAGGAACTGGACATCAACCGGCTGTCCGACTACGAT

GTGGACCACATCGTGCCTCAGAGCTTTCTGAAGGACGACTCCATCGACAACA

AGGTGCTGACCAGAAGCGACAAGGCCCGGGGCAAGAGCGACAACGTGCCCT

CCGAAGAGGTCGT GAAGAAGATGAAGA ACT ACTGGCGGC AGCTGCTGAACG

CCAAGCTGATTACCCAGAGAAAGTTCGACAATCTGACCAAGGCCGAGAGAGG

CGGCCTGAGCGAACTGGATAAGGCCGGCTTCATCAAGAGACAGCTGGTGGAA

ACCCGGCAGATCACAAAGCACGTGGCACAGATCCTGGACTCCCGGATGAACA

CTAAGTACGACGAGAATGACAAGCTGATCCGGGAAGTGAAAGTGATCACCCT

GAAGTCCAAGCTGGTGTCCGATTTCCGGAAGGATTTCCAGTTTTACAAAGTGC

GCGAGATCAACAACTACCACCACGCCCACGACGCCTACCTGAACGCCGTCGT

GGGAACCGCCCTGATCAAAAAGTACCCTAAGCTGGAAAGCGAGTTCGTGTAC

GGC G AC T AC A AGGT GT AC G AC GT GC GG A AG AT GAT C GC C A AG AGC G AGC AG

GAAATCGGCAAGGCTACCGCCAAGTACTTCTTCTACAGCAACATCATGAACTT

TTTCAAGACCGAGATTACCCTGGCCAACGGCGAGATCCGGAAGCGGCCTCTG

ATCGAGAC AAACGGCGAAACCGGGGAGATCGTGT GGGAT AAGGGCCGGGAT

TTTGCCACCGTGCGGAAAGTGCTGAGCATGCCCCAAGTGAATATCGTGAAAA

AGACCGAGGTGCAGACAGGCGGCTTCAGCAAAGAGTCTATCCTGCCCAAGAG

GAACAGCGATAAGCTGATCGCCAGAAAGAAGGACTGGGACCCTAAGAAGTA

CGGCGGCTTCGACAGCCCCACCGTGGCCTATTCTGTGCTGGTGGTGGCCAAAG

T GG A A A AGGGC A AGT C C A AG A A AC T G A AG AGT GT G A A AG AGC T GC T GGGG A

TCACCATCATGGAAAGAAGCAGCTTCGAGAAGAATCCCATCGACTTTCTGGA

AGCCAAGGGCTACAAAGAAGTGAAAAAGGACCTGATCATCAAGCTGCCTAA

GTACTCCCTGTTCGAGCTGGAAAACGGCCGGAAGAGAATGCTGGCCTCTGCC

GGCGAACTGCAGAAGGGAAACGAACTGGCCCTGCCCTCCAAATATGTGAACT

TCCTGTACCTGGCCAGCCACTATGAGAAGCTGAAGGGCTCCCCCGAGGATAA

T GAGC AG A A AC AGC T GTTT GT GGA AC AGC AC A AGC ACT ACC T GGACGAG AT C

ATCGAGCAGATCAGCGAGTTCTCCAAGAGAGTGATCCTGGCCGACGCTAATC

TGGACAAAGTGCTGTCCGCCTACAACAAGCACCGGGATAAGCCCATCAGAGA

GCAGGCCGAGAATATCATCCACCTGTTTACCCTGACCAATCTGGGAGCCCCTG

CCGCCTTCAAGTACTTTGACACCACCATCGACCGGAAGAGGTACACCAGCAC

CAAAGAGGTGCTGGACGCCACCCTGATCCACCAGAGCATCACCGGCCTGTAC

GAGACACGGATCGACCTGTCTCAGCTGGGAGGCGACagccctaagaaaaagaggaag gtg

AGCtgagaattc AAT AAAT AAAAGATCTTT ATTTTC ATT AGATCTGT GT GTT GGTTT

TTTGTGTGcggaccgagcggccgcaggaacccctagtgatggagttggccactccct ctctgcgcgctcgctcgctcac tgaggccgggcgaccaaaggtcgcccgacgcccgggctttgcccgggcggcctcagtgag cgagcgagcgcgcagctgcc tgcaggggcgcctgatgcggtattttctccttacgcatctgtgcggtatttcacaccgca tacgtcaaagcaaccatagtacgcgc cctgtagcggcgcattaagcgcggcgggtgtggtggttacgcgcagcgtgaccgctacac ttgccagcgccctagcgcccgct cctttcgctttcttcccttcctttctcgccacgttcgccggctttccccgtcaagctcta aatcgggggctccctttagggttccgattta gtgctttacggcacctcgaccccaaaaaacttgatttgggtgatggttcacgtagtgggc catcgccctgatagacggtttttcgcc ctttgacgttggagtccacgttctttaatagtggactcttgttccaaactggaacaacac tcaaccctatctcgggctattcttttgattt ataagggattttgccgatttcggcctattggttaaaaaatgagctgatttaacaaaaatt taacgcgaattttaacaaaatattaacgttt acaattttatggtgcactctcagtacaatctgctctgatgccgcatagttaagccagccc cgacacccgccaacacccgctgacgc gccctgacgggcttgtctgctcccggcatccgcttacagacaagctgtgaccgtctccgg gagctgcatgtgtcagaggttttcac cgtcatcaccgaaacgcgcgagacgaaagggcctcgtgatacgcctatttttataggtta atgtcatgataataatggtttcttagac gtcaggtggcacttttcggggaaatgtgcgcggaacccctatttgtttatttttctaaat acattcaaatatgtatccgctcatgagaca ataaccctgataaatgcttcaataatattgaaaaaggaagagtatgagtattcaacattt ccgtgtcgcccttattcccttttttgcggc attttgccttcctgtttttgctcacccagaaacgctggtgaaagtaaaagatgctgaaga tcagttgggtgcacgagtgggttacatc gaactggatctcaacagcggtaagatccttgagagttttcgccccgaagaacgttttcca atgatgagcacttttaaagttctgctat gtggcgcggtattatcccgtattgacgccgggcaagagcaactcggtcgccgcatacact attctcagaatgacttggttgagtac tcaccagtcacagaaaagcatcttacggatggcatgacagtaagagaattatgcagtgct gccataaccatgagtgataacactg cggccaacttacttctgacaacgatcggaggaccgaaggagctaaccgcttttttgcaca acatgggggatcatgtaactcgcctt gatcgttgggaaccggagctgaatgaagccataccaaacgacgagcgtgacaccacgatg cctgtagcaatggcaacaacgtt gcgcaaactattaactggcgaactacttactctagcttcccggcaacaattaatagactg gatggaggcggataaagttgcagga ccacttctgcgctcggcccttccggctggctggtttattgctgataaatctggagccggt gagcgtgggtctcgcggtatcattgca gcactggggccagatggtaagccctcccgtatcgtagttatctacacgacggggagtcag gcaactatggatgaacgaaataga cagatcgctgagataggtgcctcactgattaagcattggtaactgtcagaccaagtttac tcatatatactttagattgatttaaaactt catttttaatttaaaaggatctaggtgaagatcctttttgataatctcatgaccaaaatc ccttaacgtgagttttcgttccactgagcgt cagaccccgtagaaaagatcaaaggatcttcttgagatcctttttttctgcgcgtaatct gctgcttgcaaacaaaaaaaccaccgct accagcggtggtttgtttgccggatcaagagctaccaactctttttccgaaggtaactgg cttcagcagagcgcagataccaaata ctgtccttctagtgtagccgtagttaggccaccacttcaagaactctgtagcaccgccta catacctcgctctgctaatcctgttacc agtggctgctgccagtggcgataagtcgtgtcttaccgggttggactcaagacgatagtt accggataaggcgcagcggtcggg ctgaacggggggttcgtgcacacagcccagcttggagcgaacgacctacaccgaactgag atacctacagcgtgagctatgag aaagcgccacgcttcccgaagggagaaaggcggacaggtatccggtaagcggcagggtcg gaacaggagagcgcacgag ggagcttccagggggaaacgcctggtatctttatagtcctgtcgggtttcgccacctctg acttgagcgtcgatttttgtgatgctcg tcaggggggcggagcctatggaaaaacgccagcaacgcggcctttttacggttcctggcc ttttgctggccttttgctcacatgt pGW089-pAAV-EFSLTR-IL2-sPA (SEQ ID NO: 57)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac a tgtacagcatgcagctcgcatcctgtgtcacattgacacttgtgctccttgtcaacagcg cacccacttcaagctccacttcaagctc tacagcggaagcacagcagcagcagcagcagcagcagcagcagcagcagcacctggagca gctgttgatggacctacagg agctcctgagcaggatggagaattacaggaacctgaaactccccaggatgctcaccttca aattttacttgcccaagcaggccac agaattgaaagatcttcagtgcctagaagatgaacttggacctctgcggcatgttctgga tttgactcaaagcaaaagctttcaattg gaagatgctgagaatttcatcagcaatatcagagtaactgttgtaaaactaaagggctct gacaacacatttgagtgccaattcgat gatgagtcagcaactgtggtggactttctgaggagatggatagccttctgtcaaagcatc atctcaacaagccctcaataaGAA

TTCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTGCG

GCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCT

CGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCG

GGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATG

CGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAG

CAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGG

TTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTC

GCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTA

AATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCC

CAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAG

ACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTG

TTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATA

AGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGATTTAACAAA

AATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACT

CTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCC

AACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACA

GACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCA

TCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGG

TTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGA

AATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTAT

CCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGA

AGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCAT

TTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCT

GAAGATC AGTT GGGTGC ACGAGTGGGTT AC ATCGAACTGGATCTC AAC AGCG

GTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACT

TTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGA

GCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCAC

C AGT C AC AGA A A AGC ATCTT ACGG AT GGC AT G AC AGT A AGAGA ATT AT GC AG

TGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGA

TCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGT

AACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGAC

GAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTAT

TAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATG

GAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCT

GGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATT

GCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGA

CGGGGAGT C AGGC A ACT AT GGAT GA AC GA A AT AGAC AG AT C GCTGAGAT AG

GTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATA

CTTT AGATTGATTT AAAACTT C ATTTTT AATTT AAAAGGATCT AGGT GAAGAT

CCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTG

AGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTC

TGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGT

TTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCA

GCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCAC

CACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTT

ACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAA

GACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTG

CACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAG

CGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGG TATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCA

GGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACT

TGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAAC

GCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCAC

ATGT pGW094-pAAV EFSLTR-Ifng-P2A-Cd80-T2A-light-P2A-Gitrl-E2A-41BBL-sPA (SEQ ID NO: 58)

CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCGTCGGG

CGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGT

GGCCAACTCCATCACTAGGGGTTCCTGCGGCCTCTAGAAAGGATCTGCGATC

GCTCCGGTGCCCGTCAGTGGGCAGAGCGCACATCGCCCACAGTCCCCGAGAA

GTTGGGGGGAGGGGTCGGC A ATT GAACGGGTGCCT AGAGAAGGT GGCGCGG

GGTAAACTGGGAAAGTGATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTG

GGGGAGAACCGTATATAAGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAAC

GGGTTTGCCGCCAGAACACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTC

ACGCGCCCGCCGCCCTACCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTC

TGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGT

TTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACC

TAGACTCAGCCGGCTCTCCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACG

TCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAGATCTGGCTAACTAccggtgccac c atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgcGGATCCGGCGCAACAAACTTCTCTCTGC TGAA

ACAAGCCGGAGATGTCGAAGAGAATCCTGGACCGatggcttgcaattgtcagttgat gcaggat acaccactcctcaagtttccatgtccaaggctcattcttctctttgtgctgctgattcgt ctttcacaagtgtcttcagatgttgatgaac aactgtccaagtcagtgaaagataaggtattgctgccttgccgttacaactctcctcatg aagatgagtctgaagaccgaatctact ggcaaaaacatgacaaagtggtgctgtctgtcattgTtgggaaactaaaagtgtggcccg agtataagaaccggactttatatga caacactacctactctcttatcatcctgggcctggtcctttcagaccggggcacatacag ctgtgtcgttcaaaagaaggaaagag gaacgtatgaagttaaacacttggctttagtaaagttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaaccc atctgcagacactaaaaggattacctgctttgcttccgggggtttcccaaagcctcgctt ctcttggttggaaaatggaagagaatta cctggcatcaatacgacaatttcccaggatcctgaatctgaattgtacaccattagtagc caactagatttcaatacgactcgcaac cacaccattaagtgtctcattaaatatggagatgctcacgtgtcagaggacttcacctgg gaaaaacccccagaagaccctcctg atagcaagaacacacttgtgctctttggggcaggattcggcgcagtaataacagtcgtcg tcatcgttgtcatcatcaaatgcttctg taagcacagaagctgtttcagaagaaatgaggcaagcagagaaacaaacaacagccttac cttcgggcctgaagaagcattag ctgaacagaccgtcttccttcgtacgGGC AGTGGAGAGGGC AGAGGAAGTCTGCT AAC AT GC

GGTGACGTCGAGGAGAATCCTGGCCCAatggagagtgtggtacagccttcagtgttt gtggtggatgga cagacggacatcccattcaggcggctggaacagaaccaccggagacggcgctgtggcact gtccaggtcagcctggccctgg tgctgctgctaggtgctgggctggccactcagggctggtttctcctgagactgcatcaac gtcttggagacatagtagctcatctgc cagatggaggcaaaggctcctgggagaagctgatacaagatcaacgatctcaccaggcca acccagcagcacatcttacagg agccaacgccagcttgataggtattggtggacctctgttatgggagacacgacttggcct ggccttcttgaggggcttgacgtatc atgatggggccctggtgaccatggagcccggttactactatgtgtactccaaagtgcagc tgagcggcgtgggctgcccccagg ggctggccaatggcctccccatcacccatggactatacaagcgcacatcccgctacccga aggagttagaactgctggtcagtc ggcggtcaccctgtggccgggccaacagctcccgagtctggtgggacagcagcttcctgg gcggcgtggtacatctggaggc tggggaagaggtggtggtccgcgtgcctggaaaccgcctggtcagaccacgtgacggcac caggtcctatttcggagctttcat ggtcactagtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATG

TCGAAGAGAATCCTGGACCGatggaggaaatgcctttgagagagtcaagtcctcaaa gggcagagaggtgc aagaagtcatggctcttgtgcatagtggctctgttactgatgctgctctgttctttgggt acactgatctatacttcactcaagccaact gccatcgagtcctgcatggttaagtttgaactatcatcctcaaaatggcacatgacatct cccaaacctcactgtgtgaatacgacat ctgatgggaagctgaagatactgcagagtggcacatatttaatctacggccaagtgattc ctgtggataagaaatacataaaagac aatgcccccttcgtagtacagatatataaaaagaatgatgtcctacaaactctaatgaat gattttcaaatcttgcctataggaggggt ttatgaactgcatgctggagataacatatatctgaagttcaactctaaagaccatattca gaaaaataacacatactgggggatcatc ttaatgcctgatctaccattcatctctgctagcagcggtacccagtgcaccaactacgcc ctgctgaagctggccggcgatgtgga gagcaaccccgggcccatggaccagcacacacttgatgtggaggataccgcggatgccag acatccagcaggtacttcgtgc ccctcggatgcggcgctcctcagagataccgggctcctcgcggacgctgcgctcctctca gatactgtgcgccccacaaatgcc gcgctccccacggatgctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtgg ccgcctgcactgaacttctgttccc gccacccaaagctctatggcctagtcgctttggttttgctgcttctgatcgccgcctgtg ttcctatcttcacccgcaccgagcctcg gccagcgctcacaatcaccacctcgcccaacctgggtacccgagagaataatgcagacca ggtcacccctgtttcccacattgg ctgccccaacactacacaacagggctctcctgtgttcgccaagctactggctaaaaacca agcatcgttgtgcaatacaactctga actggcacagccaagatggagctgggagctcatacctatctcaaggtctgaggtacgaag aagacaaaaaggagttggtggta gacagtcccgggctctactacgtatttttggaactgaagctcagtccaacattcacaaac acaggccacaaggtgcagggctgg gtctctcttgttttgcaagcaaagcctcaggtagatgactttgacaacttggccctgaca gtggaactgttcccttgctccatggaga acaagttagtggaccgttcctggagtcaactgttgctcctgaaggctggccaccgcctca gtgtgggtctgagggcttatctgcat ggagcccaggatgcatacagagactgggagctgtcttatcccaacaccaccagctttgga ctctttcttgtgaaacccgacaacc catgggaatgaGAATTCAATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTT

TTTGTGTGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTG

CGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGG

GCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGG

GCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCAT

ACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGG

GTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCTTAGCGCCC

GCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTC

AAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCAC

CTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCC

CTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTG

GACTCTTGTTCCAAACTGGAACAACACTCAACTCTATCTCGGGCTATTCTTTT

GATTTATAAGGGATTTTGCCGATTTCGGTCTATTGGTTAAAAAATGAGCTGAT

TTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTAT

GGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGA

CACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATC

CGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTT

CACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATT

TTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTT

TTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCA

AATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTG

AAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTT

TTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTA

A A AG AT GC T GA AG AT C AGTT GGGT GC AC GAGT GGGTT AC ATCGA ACTGGAT C

TCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATG

ATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGC

CGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTT

GAGT ACTC AC C AGT C AC AGA A A AGC AT C TT ACGGAT GGC AT GAC AGT A AGAG

AATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTT

CTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGG

GGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCAT

ACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTG

CGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAAT AGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTT

CCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCC

GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTT

ATCT AC ACGACGGGGAGT C AGGC AACT AT GGAT GAACGAA AT AGAC AGATCG

CTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTAC

T CAT AT AT ACTTT AGATT GATTT AAAACTT C ATTTTT AATTT AAAAGGATCT AG

GTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTC

GTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGAT

CCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACC

AGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAA

CTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAG

TTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCT

AATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGT

TGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGG

GGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGA

TACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGG

CGGAC AGGT ATCCGGT AAGCGGC AGGGTCGGAAC AGGAGAGCGC ACGAGGG

AGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCAC

CTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATG

GAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTT

TTGCTCACATGT

Ori (SEQ ID NO: 59)

TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCAC

CGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCG

AAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTA

GCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCG

CTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTT

ACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCT

GAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGA

ACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGG

AGAAAGGCGGAC AGGT ATCCGGTAAGCGGC AGGGTCGGAAC AGGAGAGCGC

ACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTT

TCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGA

GCCTATGGAAA flOri (SEQ ID NO: 60)

ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAG

CGTGACCGCTACACTTGCCAGCGCCTTAGCGCCCGCTCCTTTCGCTTTCTTCCC

TTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCT

CCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTG

ATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGC

CCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGG

AACAACACTCAACTCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCC

GATTTCGGTCT ATT GGTT AAAAAAT GAGCTGATTT AAC AAAAATTT AACGCGA

ATTTTAACAAAAT ATT AACGTTTAC AATTT

AmpR Promoter (SEQ ID NO: 61)

CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTC AT GAGAC A AT A AC CC T GAT A A AT GC TT C A AT A AT ATT GA A A A AGG A AGAGT AmpR (SEQ ID NO: 62)

ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGC

CTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGA

TCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAG

ATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAA

AGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAAC

TCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTC

AC AGAAAAGC ATCTT ACGGAT GGC AT GAC AGT AAGAGAATT AT GC AGT GCTG

CCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGG

AGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACT

CGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGC

GTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAAC

TGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAG

GCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTT

TATTGCTGATAAATCTGGAGCCGGTGAGCGTGGAAGCCGCGGTATCATTGCA

GCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGG

GG AGT C AGGC A AC TAT GG AT G A AC G A A AT AG AC AG AT C GC T GAG AT AGGT GC

CTCACTGATTAAGCATTGGTAA

AAV-ITR (SEQ ID NO: 63)

AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTC

ACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGG

CCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG

EFS core promoter (SEQ ID NO: 64)

GGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCG

GCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTG

ATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATA

AGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAAC

ACAG

LTR promoter (SEQ ID NO: 65)

GGGGCTCGCATCTCTCCTTCACGCGCCCGCCGCCCTACCTGAGGCCGCCATCC

ACGCCGGTTGAGTCGCGTTCTGCCGCCTCCCGCCTGTGGTGCCTCCTGAACTG

CGTCCGCCGTCTAGGTAAGTTTAAAGCTCAGGTCGAGACCGGGCCTTTGTCCG

GCGCTCCCTTGGAGCCTACCTAGACTCAGCCGGCTCTCCACGCTTTGCCTGAC

CCTGCTTGCTCAACTCTACGTCTTTGTTTCGTTTTCTGTTCTGCGCCGTTACAG

ATC

EFS-LTR promoter (SEQ ID NO: 66)

GGGCAGAGCGCACATCGCCCACAGTCCCCGAGAAGTTGGGGGGAGGGGTCG

GCAATTGAACGGGTGCCTAGAGAAGGTGGCGCGGGGTAAACTGGGAAAGTG

ATGTCGTGTACTGGCTCCGCCTTTTTCCCGAGGGTGGGGGAGAACCGTATATA

AGTGCAGTAGTCGCCGTGAACGTTCTTTTTCGCAACGGGTTTGCCGCCAGAAC

ACAGCTGAAGCTTCGAGGGGCTCGCATCTCTCCTTCACGCGCCCGCCGCCCTA

CCTGAGGCCGCCATCCACGCCGGTTGAGTCGCGTTCTGCCGCCTCCCGCCTGT

GGTGCCTCCTGAACTGCGTCCGCCGTCTAGGTAAGTTTAAAGCTCAGGTCGAG

ACCGGGCCTTTGTCCGGCGCTCCCTTGGAGCCTACCTAGACTCAGCCGGCTCT CCACGCTTTGCCTGACCCTGCTTGCTCAACTCTACGTCTTTGTTTCGTTTTCTG

TTCTGCGCCGTTACAGATC

T2A (SEQ ID NO: 67)

GAGGA AGT C T GC T A AC AT GC GGT GACGTCGAGGAG A AT C CTGGCC C Aatggagag tgtggtacagcct

P2A (SEQ ID NO: 68) actagtGGATCCGGCGCAACAAACTTCTCTCTGCTGAAACAAGCCGGAGATGTCG

AAGAGAATCCTGGACCG

E2A (SEQ ID NO: 69) gctagcagcggtacccagtgcaccaactacgccctgctgaagctggccggcgatgtggag agcaaccccgggccc

F2A (SEQ ID NO: 70) ggcatatgcggtaccgtgaagcagaccctgaacttcgatctgctgaagctggccggcgat gtggagagcaaccccgggccc

WPRE (SEQ ID NO: 71) aatcaacctctggattacaaaatttgtgaaagattgactggtattcttaactatgttgct ccttttacgctatgtggatacgctgctttaat gcctttgtatcatgctattgcttcccgtatggctttcattttctcctccttgtataaatc ctggttgctgtctctttatgaggagttgtggcc cgttgtcaggcaacgtggcgtggtgtgcactgtgtttgctgacgcaacccccactggttg gggcattgccaccacctgtcagctc ctttccgggactttcgctttccccctccctattgccacggcggaactcatcgccgcctgc cttgcccgctgctggacaggggctcg gctgttgggcactgacaattccgtggtgttgtcggggaaatcatcgtcctttccttggct gctcgcctgtgttgccacctggattctg cgcgggacgtccttctgctacgtcccttcggccctcaatccagcggaccttccttcccgc ggcctgctgccggctctgcggcctct tccgcgtcttcgccttcgccctcagacgagtcggatctccctttgggccgcctccccgc short-PolyA (SEQ ID NO: 72)

AATAAAAGATCTTTATTTTCATTAGATCTGTGTGTTGGTTTTTTGTGTG

ORF-Cd80 (SEQ ID NO: 73) atggcttgcaattgtcagttgatgcaggatacaccactcctcaagtttccatgtccaagg ctcattcttctctttgtgctgctgattcgtc tttcacaagtgtcttcagatgttgatgaacaactgtccaagtcagtgaaagataaggtat tgctgccttgccgttacaactctcctcat gaagatgagtctgaagaccgaatctactggcaaaaacatgacaaagtggtgctgtctgtc attgctgggaaactaaaagtgtggc ccgagtataagaaccggactttatatgacaacactacctactctcttatcatcctgggcc tggtcctttcagaccggggcacataca gctgtgtcgttcaaaagaaggaaagaggaacgtatgaagttaaacacttggctttagtaa agttgtccatcaaagctgacttctcta cccccaacataactgagtctggaaacccatctgcagacactaaaaggattacctgctttg cttccgggggtttcccaaagcctcgc ttctcttggttggaaaatggaagagaattacctggcatcaatacgacaatttcccaggat cctgaatctgaattgtacaccattagta gccaactagatttcaatacgactcgcaaccacaccattaagtgtctcattaaatatggag atgctcacgtgtcagaggacttcacct gggaaaaacccccagaagaccctcctgatagcaagaacacacttgtgctctttggggcag gattcggcgcagtaataacagtcg tcgtcatcgttgtcatcatcaaatgcttctgtaagcacagaagctgtttcagaagaaatg aggcaagcagagaaacaaacaacag ccttaccttcgggcctgaagaagcattagctgaacagaccgtcttcctt

ORF-Light (SEQ ID NO: 74) atggagagtgtggtacagccttcagtgtttgtggtggatggacagacggacatcccattc aggcggctggaacagaaccaccgg agacggcgctgtggcactgtccaggtcagcctggccctggtgctgctgctaggtgctggg ctggccactcagggctggtttctc ctgagactgcatcaacgtcttggagacatagtagctcatctgccagatggaggcaaaggc tcctgggagaagctgatacaagat caacgatctcaccaggccaacccagcagcacatcttacaggagccaacgccagcttgata ggtattggtggacctctgttatggg agacacgacttggcctggccttcttgaggggcttgacgtatcatgatggggccctggtga ccatggagcccggttactactatgtg tactccaaagtgcagctgagcggcgtgggctgcccccaggggctggccaatggcctcccc atcacccatggactatacaagcg cacatcccgctacccgaaggagttagaactgctggtcagtcggcggtcaccctgtggccg ggccaacagctcccgagtctggt gggacagcagcttcctgggcggcgtggtacatctggaggctggggaagaggtggtggtcc gcgtgcctggaaaccgcctggt cagaccacgtgacggcaccaggtcctatttcggagctttcatggtc

ORF-CxcllO (SEQ ID NO: 75) atgaacccaagtgctgccgtcattttctgcctcatcctgctgggtctgagtgggactcaa gggatccctctcgcaaggacggtccg ctgcaactgcatccatatcgatgacgggccagtgagaatgagggccatagggaagcttga aatcatccctgcgagcctatcctg cccacgtgttgagatcattgccacgatgaaaaagaatgatgagcagagatgtctgaatcc ggaatctaagaccatcaagaatttaa tgaaagcgtttagccaaaaaaggtctaaaagggctcct

ORF-41BBL (SEQ ID NO: 76) atggaccagcacacacttgatgtggaggataccgcggatgccagacatccagcaggtact tcgtgcccctcggatgcggcgct cctcagagataccgggctcctcgcggacgctgcgctcctctcagatactgtgcgccccac aaatgccgcgctccccacggatg ctgcctaccctgcggttaatgttcgggatcgcgaggccgcgtggccgcctgcactgaact tctgttcccgccacccaaagctctat ggcctagtcgctttggttttgctgcttctgatcgccgcctgtgttcctatcttcacccgc accgagcctcggccagcgctcacaatca ccacctcgcccaacctgggtacccgagagaataatgcagaccaggtcacccctgtttccc acattggctgccccaacactacac aacagggctctcctgtgttcgccaagctactggctaaaaaccaagcatcgttgtgcaata caactctgaactggcacagccaaga tggagctgggagctcatacctatctcaaggtctgaggtacgaagaagacaaaaaggagtt ggtggtagacagtcccgggctcta ctacgtatttttggaactgaagctcagtccaacattcacaaacacaggccacaaggtgca gggctgggtctctcttgttttgcaagc aaagcctcaggtagatgactttgacaacttggccctgacagtggaactgttcccttgctc catggagaacaagttagtggaccgtt cctggagtcaactgttgctcctgaaggctggccaccgcctcagtgtgggtctgagggctt atctgcatggagcccaggatgcata cagagactgggagctgtcttatcccaacaccaccagctttggactctttcttAtgaaacc cgacaacccatgggaa

ORF-IFNg (SEQ ID NO: 77) atgaacgctacacactgcatcttggctttgcagctcttcctcatggctgtttctggctgt tactgccacggcacagtcattgaaagcct agaaagtctgaataactattttaactcaagtggcatagatgtggaagaaaagagtctctt cttggatatctggaggaactggcaaaa ggatggtgacatgaaaatcctgcagagccagattatctctttctacctcagactctttga agtcttgaaagacaatcaggccatcag caacaacataagcgtcattgaatcacacctgattactaccttcttcagcaacagcaaggc gaaaaaggatgcattcatgagtattg ccaagtttgaggtcaacaacccacaggtccagcgccaagcattcaatgagctcatccgag tggtccaccagctgttgccggaat ccagcctcaggaagcggaaaaggagtcgctgctga

ORF-I12 (SEQ ID NO: 78) atgtacagcatgcagctcgcatcctgtgtcacattgacacttgtgctccttgtcaacagc gcacccacttcaagctccacttcaagct ctacagcggaagcacagcagcagcagcagcagcagcagcagcagcagcagcacctggagc agctgttgatggacctacag gagctcctgagcaggatggagaattacaggaacctgaaactccccaggatgctcaccttc aaattttacttgcccaagcaggcca cagaattgaaagatcttcagtgcctagaagatgaacttggacctctgcggcatgttctgg atttgactcaaagcaaaagctttcaatt ggaagatgctgagaatttcatcagcaatatcagagtaactgttgtaaaactaaagggctc tgacaacacatttgagtgccaattcga tgatgagtcagcaactgtggtggactttctgaggagatggatagccttctgtcaaagcat catctcaacaagccctcaataaGA

ATTC

ORF-Gitrl (SEQ ID NO: 79) atggaggaaatgcctttgagagagtcaagtcctcaaagggcagagaggtgcaagaagtca tggctcttgtgcatagtggctctgt tactgatgctgctctgttctttgggtacactgatctatacttcactcaagccaactgcca tcgagtcctgcatggttaagtttgaactat catcctcaaaatggcacatgacatctcccaaacctcactgtgtgaatacgacatctgatg ggaagctgaagatactgcagagtgg cacatatttaatctacggccaagtgattcctgtggataagaaatacataaaagacaatgc ccccttcgtagtacagatatataaaaa gaatgatgtcctacaaactctaatgaatgattttcaaatcttgcctataggaggggttta tgaactgcatgctggagataacatatatc tgaagttcaactctaaagaccatattcagaaaaataacacatactgggggatcatcttaa tgcctgatctaccattcatctct

ORF-I123 (SEQ ID NO: 80)

ATGTGTCCTCAGAAGCTAACCATCTCCTGGTTTGCCATCGTTTTGCTGGTGTCT

CCACTCATGGCCATGTGGGAGCTGGAGAAAGACGTTTATGTTGTAGAGGTGG ACTGGACTCCCGATGCCCCTGGAGAAACAGTGAACCTCACCTGTGACACGCC

TGAAGAAGATGACATCACCTGGACCTCAGACCAGAGACATGGAGTCATAGGC

TCTGGAAAGACCCTGACCATCACTGTCAAAGAGTTTCTAGATGCTGGCCAGTA

CACCTGCCACAAAGGAGGCGAGACTCTGAGCCACTCACATCTGCTGCTCCAC

AAGAAGGAAAAT GGAATTT GGTCC ACTGAAATTTT AAAAAATTT C AAAA AC A

AGACTTTCCTGAAGTGTGAAGCACCAAATTACTCCGGACGGTTCACGTGCTCA

TGGCTGGTGCAAAGAAACATGGACTTGAAGTTCAACATCAAGAGCAGTAGCA

GTTCCCCTGACTCTCGGGCAGTGACATGTGGAATGGCGTCTCTGTCTGCAGAG

AAGGTCACACTGGACCAAAGGGACTATGAGAAGTATTCAGTGTCCTGCCAGG

AGGATGTCACCTGCCCAACTGCCGAGGAGACCCTGCCCATTGAACTGGCGTT

GGAAGCACGGCAGCAGAATAAATATGAGAACTACAGCACCAGCTTCTTCATC

AGGGACATCATCAAACCAGACCCGCCCAAGAACTTGCAGATGAAGCCTTTGA

AGAACTCACAGGTGGAGGTCAGCTGGGAGTACCCTGACTCCTGGAGCACTCC

CCATTCCTACTTCTCCCTCAAGTTCTTTGTTCGAATCCAGCGCAAGAAAGAAA

AG AT G A AGG AG AC AG AGG AGGGGT GT A AC C AG A A AGGT GC GT TC C T C GT AG

AGAAGAC ATCT ACCGAAGTCC AATGC AAAGGCGGGAAT GTCTGCGT GC AAGC

T C AGGATCGCT ATT AC A ATTCCTC AT GC AGC AAGT GGGC AT GT GTTCCCTGC A

GGGTCCGATCCTAG

ORF-hIFNg (SEQ ID NO: 81)

ATGAAATATACAAGTTATATCTTGGCTTTTCAGCTCTGCATCGTTTTGGGTTCT

CTTGGCTGTTACTGCCAGGACCCATATGTAAAAGAAGCAGAAAACCTTAAGA

AATATTTTAATGCAGGTCATTCAGATGTAGCGGATAATGGAACTCTTTTCTTA

GGCATTTTGAAGAATTGGAAAGAGGAGAGTGACAGAAAAATAATGCAGAGC

CAAATTGTCTCCTTTTACTTCAAACTTTTTAAAAACTTTAAAGATGACCAGAG

C ATCC AAAAGAGT GT GGAGACC ATC AAGGAAGAC AT GAAT GTC AAGTTTTT C

AATAGCAACAAAAAGAAACGAGATGACTTCGAAAAGCTGACTAATTATTCGG

T AACTGACTT GAAT GTCC A AC GCA A AGC A AT AC AT GAACTC ATCC AAGT GAT

GGCTGAACTGTCGCCAGCAGCTAAAACAGGGAAGCGAAAAAGGAGTCAGAT

GCTGTTTCGAGGTCGAAGAGCATCCCAG

ORF-hIL23 (SEQ ID NO: 82)

TGTGTCACCAGCAGTTGGTCATCTCTTGGTTTTCCCTGGTTTTTCTGGCATCTC

CCCTCGT GGCC AT AT GGGAACTGAAGA AAGAT GTTT AT GTCGT AGAATT GGA

TTGGTATCCGGATGCCCCTGGAGAAATGGTGGTCCTCACCTGTGACACCCCTG

AAGAAGATGGTATCACCTGGACCTTGGACCAGAGCAGTGAGGTCTTAGGCTC

TGGCAAAACCCTGACCATCCAAGTCAAAGAGTTTGGAGATGCTGGCCAGTAC

ACCTGTCACAAAGGAGGCGAGGTTCTAAGCCATTCGCTCCTGCTGCTTCACAA

A A AGG A AG AT GGAATTT GGT C C ACTGAT ATTTT A A AGG AC C AG A A AG A ACC C

AAAAATAAGACCTTTCTAAGATGCGAGGCCAAGAATTATTCTGGACGTTTCA

CCTGCTGGTGGCTGACGACAATCAGTACTGATTTGACATTCAGTGTCAAAAGC

AGCAGAGGCTCTTCTGACCCCCAAGGGGTGACGTGCGGAGCTGCTACACTCT

CTGC AG AGAGAGT C AGAGGGGAC A AC A AGGAGT AT GAGT AC TC AGT GG AGT

GCCAGGAGGACAGTGCCTGCCCAGCTGCTGAGGAGAGTCTGCCCATTGAGGT

CATGGTGGATGCCGTTCACAAGCTCAAGTATGAAAACTACACCAGCAGCTTC

TTCATCAGGGACATCATCAAACCTGACCCACCCAAGAACTTGCAGCTGAAGC

CATTAAAGAATTCTCGGCAGGTGGAGGTCAGCTGGGAGTACCCTGACACCTG

GAGTACTCCACATTCCTACTTCTCCCTGACATTCTGCGTTCAGGTCCAGGGCA

AGAGCAAGAGAGAAAAGAAAGATAGAGTCTTCACGGACAAGACCTCAGCCA CGGTCATCTGCCGCAAAAATGCCAGCATTAGCGTGCGGGCCCAGGACCGCTA

CTATAGCTCATCTTGGAGCGAATGGGCATCTGTGCCCTGCAGT

ORF-h41BBL (SEQ ID NO: 83)

ATGGAATACGCCTCTGACGCTTCACTGGACCCCGAAGCCCCGTGGCCTCCCGC

GCCCCGCGCTCGCGCCTGCCGCGTACTGCCTTGGGCCCTGGTCGCGGGGCTGC

TGCTGCTGCTGCTGCTCGCTGCCGCCTGCGCCGTCTTCCTCGCCTGCCCCTGG

GCCGTGTCCGGGGCTCGCGCCTCGCCCGGCTCCGCGGCCAGCCCGAGACTCC

GCGAGGGTCCCGAGCTTTCGCCCGACGATCCCGCCGGCCTCTTGGACCTGCG

GCAGGGCATGTTTGCGCAGCTGGTGGCCCAAAATGTTCTGCTGATCGATGGG

CCCCTGAGCTGGTACAGTGACCCAGGCCTGGCAGGCGTGTCCCTGACGGGGG

GCCTGAGCTACAAAGAGGACACGAAGGAGCTGGTGGTGGCCAAGGCTGGAG

TCTACTATGTCTTCTTTCAACTAGAGCTGCGGCGCGTGGTGGCCGGCGAGGGC

TCAGGCTCCGTTTCACTTGCGCTGCACCTGCAGCCACTGCGCTCTGCTGCTGG

GGCCGCCGCCCTGGCTTTGACCGTGGACCTGCCACCCGCCTCCTCCGAGGCTC

GGAACTCGGCCTTCGGTTTCCAGGGCCGCTTGCTGCACCTGAGTGCCGGCCAG

CGCCTGGGCGTCCATCTTCACACTGAGGCCAGGGCACGCCATGCCTGGCAGC

TTACCCAGGGCGCCACAGTCTTGGGACTCTTCCGGGTGACCCCCGAAATCCCA

GCCGGACTCCCTTCACCGAGGTCGGAA

ORF-hLIGHT(remove-EQLI) (SEQ ID NO: 84)

ATGGAGGAGAGTGTCGTACGGCCCTCAGTGTTTGTGGTGGATGGACAGACCG

ACATCCCATTCACGAGGCTGGGACGAAGCCACCGGAGACAGTCGTGCAGTGT

GGCCCGGGTGGGTCTGGGTCTCTTGCTGTTGCTGATGGGGGCCGGGCTGGCCG

TCCAAGGCTGGTTCCTCCTGCAGCTGCACTGGCGTCTAGGAGAGATGGTCACC

CGCCTGCCTGACGGACCTGCAGGCTCCTGGCAAGAGCGAAGGTCTCACGAGG

TCAACCCAGCAGCGCATCTCACAGGGGCCAACTCCAGCTTGACCGGCAGCGG

GGGGCCGCTGTTATGGGAGACTCAGCTGGGCCTGGCCTTCCTGAGGGGCCTC

AGCTACCACGATGGGGCCCTTGTGGTCACCAAAGCTGGCTACTACTACATCTA

CTCCAAGGTGCAGCTGGGCGGTGTGGGCTGCCCGCTGGGCCTGGCCAGCACC

ATCACCCACGGCCTCTACAAGCGCACACCCCGCTACCCCGAGGAGCTGGAGC

TGTTGGTCAGCCAGCAGTCACCCTGCGGACGGGCCACCAGCAGCTCCCGGGT

CTGGT GGGAC AGC AGCTTCCTGGGTGGT GT GGT AC ACCTGGAGGCTGGGGAG

AAGGTGGTCGTCCGTGTGCTGGATGAACGCCTGGTTCGACTGCGTGATGGTAC

CCGGTCTTACTTCGGGGCTTTCATGGTG

CGKRK (SEQ ID NO: 85) tgcggaaagcgtaag

The results of the experiments are now described.

Example 1: CRISPRa screen assisted rational design of an off the shelf viral immune gene therapy

In order to develop an improved off-the-shelf viral immune gene therapy that is both simpler and more potent, an in vivo CRISPRa-based screen was devised using a sgRNA library to identify the key genes that are effective in Multiplexed Activation of Endogenous Genes as an Immunotherapy (MAEGI) or direct ORF -based viral immune gene therapy settings (Fig. 1 A). By performing a CRISPRa in vivo depletion screen in the syngeneic systemic cancer model, a small number of genes that are depleted in the metastatic tumors in the lung were identified.

An APCM sgRNA library (SEQ ID NOs. 86-192) (Fig. 7) was introduced into E0771, a triple-negative breast cancer cell line, by infection with a lentiviral CRISPRa vector at M.O.I = 0.2-0.3. After 7 days of in vitro culture under puromycin, 2*10 ⁶ of APCM library transduced cells were intravenously injected into immune-competent C57BL/6J mice (Fig. 1 A). Mouse survival was then followed for 30 days (Fig. IB). The metastatic tumors formed in the lungs of each animal were collected, and the immune- mediated sgRNA depletion and enrichment was readout by illumina sequencing (Fig.

IC). The abundance of sgRNAs within tumor samples and cell samples was then determined by mapping the sequenced reads to the library. The depleted genes were chosen in combinations (e.g. CLC4, CLC4G, CLC4I) as screen-rationalized, off-the-shelf viral immune gene therapy, either in the form of o-MAEGI or concatenated ORF (Fig.

ID).

Example 2: In vivo anti-tumor efficacy of AAV-0-MAEGI-CLC4. AAV-ORF-CLC4 and AAV-ORF-CLC4I in a syngeneic triple-negative breast cancer model

Taking the top hits from the analysis of depleted gene expression in APCM- transduced tumors from Example 1, three off-the-shelf therapies were developed: CLC4 (CD80-Light-CXCL 10-4- 1BBL), CLC4G (CD80-Light-CXCL10-4-lBBL-Gitrl) and CLC4I (CD80-Light-CXCL10-4-lBBL-IFNG). These therapies were developed in the form of MAEGI (CRISPRa-based pooled activation) and direct ORF -based viral immune gene therapy (AAV expressing concatenated ORFs).

The therapeutic effects of intratumoral activation of CLC4, CLC4G and CLC4I were assessed by injecting the AAV constructs into established E077 tumors. Mice were treated with PBS, AAV-dCas9 + Vector, AAV-dCas9 + CLC4 (dual AAV delivered CRISPRa of sgRNA library targeting CLC4G, i.e. 0-MAEGI-CLC4G), AAV-orf-CLC4 (AAV delivered open reading frame expression of CD80-Light-CXCL10-4-lBBL), or AAV-orf-CLC4I (AAV delivered open reading frame expression of CD80-Light- CXCL10-4-1BBL-IFNG). Tumor growth was then followed over the next 27 days (Fig. 2). Tumors treated with empty vector plus the dCas9 AAV (Vector + 1 lb) had slightly reduced but not significant growth compared to PBS-alone controls. Treatment with CLC4 (pGW066) resulted in a significant reduction in tumor volume. Treatment with CLC4G+1 lb (o-MAEGI) or CLC4I (pGW067) resulted in a highly significant reduction in tumor volume. The data demonstrated the in vivo therapeutic efficacy of these treatments.

Example 3: Design and characterization of adenovirus-based one-vector CRISPRa systems

Adenoviral-based dSpCas9 CRISPRa systems were designed herein that utilize a single construct to deliver all CRISPRa components. The constructs, pGW029 (SEQ ID NO: 4), pGW035 (SEQ ID NO: 5), and pGW063 (SEQ ID NO: 6), are illustrated in Fig.

3 A. In order to test this system in vitro, an sgRNA that targed Ox40L was cloned into these constructs, and their activation efficacies were tested. Four days post-transfection, expression levels of Ox40L were determined by quantitative RT-PCR (Fig. 3B). The cytotoxic effects in HEK293FT cells caused by adenovirus production were then observed (Fig. 3C). Cells were transfected with AdTrack (adenoviral CMV-EGFP) constructs, and the expression of GFP and cytopathic effects were observed (Fig. 3C top row). In a similar study, the CRISPRa system was cloned into adenoviral constructs (U6- sgRNA-EFS-LTR-dSpCas9-p65-HSFl-sPA) and the adenoviruses were rescued. 4-6 days post-infection of the adenovirus, cytopathic effects on HEK293FT were observed (Fig.

3C, bottom row). These results demonstrated the robustness of the dSpCas9 CRISPRa adenoviral system.

Example 4: Design and testing of CRISPRa AAV-based vector systems

Additional vector systems were designed utilizing AAV to deliver CRISPRa components to target cells. The first was a dSaCas9-based AIO (all-in-one) CRISPRa system, which uses a single AAV vector (pZB3: SEQ ID NO: 10) to deliver all CRISPRa components (Fig. 4A). For in vitro testing of this system, Pmel was targeted with the AIO CRISPRa system. Four days post-infection, expression levels of Pmel were determined by quantitative RT-PCR, which revealed up-regulation of Pmel expression by the AAV- based (pZB3) vector (Fig. 4B). A lentiviral based version was used for comparison.

An AAV-based two-vector system with dSaCas9 was designed for use in endogenous gene activation and MAEGI. In this case, the dSaCas9 enzyme was delivered on a separate construct from the other CRISPRa components (pGW060: SEQ ID NO: 12 and pGW047: SEQ ID NO: 11, respectively) (Fig. 5A). For in vitro testing, Pmel targeted sgRNA was cloned into the dual AAV-CRISPRa system. Expression levels of Pmel were determined by quantitative RT-PCR four days after infection (Fig. 5B). A significant increase in Pmel expression was observed in cells transfected with the two-vector dSaCas9 system.

Another two-vector system was also designed, which utilized the dSpCas9 endonuclease. A schematic of the two constructs used in this dSpCas9-based dual-AAV delivery system is depicted in Fig. 6A (pGW045: SEQ ID NO: 13). For in vitro testing, Pmel targeted sgRNA was cloned into cells with the dual AAV-CRISPRa system (Fig. 6B). Four days post-infection, expression levels of Pmel were determined by quantitative RT-PCR, which demonstrated significant upregulation of Pmel transcript as compared to a non-sgRNA containing control.

Example 5: An AAV-based two-vector system with dCas!2a/dCpfl

AAV-based two-vector systems were generated that utilize dCasl2a/dCpfl to facilitate endogenous gene activation and MAEGI. Vectors comprising Casl2 (formerly known as Cpfl) derived from Lachnospiraceae bacterium ND2006 (dLbCasl2a/ dLbCpfl) or A cidaminococcus sp. (dAsCasl2a/dAsCpfl) were generated. Vectors used in this system include: pRCl 19: pRCl 19 AAV EFS-denAsCasl2a-SunTag3x-l (SEQ ID NO: 14); pRC120: pRC120 AAV EFS-dLbCasl2a-SunTag5x-l (SEQ ID NO: 15); pRC121b: pRC121b AAV-EFS-LTR-scFV_Gcn4-Activ-Triplex-SapI-WPRE (SEQ ID NO: 16); pRC124: pRC124 AAV-EFS-LTR-NLS-dLbCpfl_574-NLS-Activ-Triplex- SapI-WPRE-1 (SEQ ID NO: 17); and pRC126: pRC126 AAV -EF S -LTR-NL S - Acti v- dLbCpfl_1229-NLS-Activ-sPA-l(SEQ ID NO: 18).

Example 6: Multiplexed activation of endogenous genes by APCM as an immune-gene therapy approach

Syngeneic orthotopic breast tumors were induced in mice by E0771 cells in C57BL/6 mice (Figs. 9A-9D). These animals were treated with PBS, AAV-Vector, or AAV-APCM sgRNA library by intratumoral administration at indicated times (arrows), using either a single vector carrying the library alone into dCas9-VP64 expressing cells (Fig. 9A, against single tumor; and Fig.9C, against both local and distant tumor), or using a dual-AAV approach (Fig. 9B). Survival of these animals treated with PBS, AAV- Vector, or AAV-APCM sgRNA library showed that AAV-APCM treatment extended the overall survival of treated animals (Fig. 9D). Syngeneic pancreatic tumors were induced in mice by Pan02 cells in C57BL/6 mice (Fig. 9E). These animals were treated with PBS, AAV- Vector, AAV-p-MAEGI(Pan02) or AAV-APCM sgRNA library by intratumoral administration with two doses per week, total of 3-4 doses starting at end of week 1.

Example 7: Molecular characterization and therapeutic effect of multiplexed activation of Cd80. Light CxcllO. Gitrh 41bbh and IFNg in tumor cells by AAV-delivered CRISPRa or AAV-ORFs (polvcistronic open-reading-frames)

Fig. 10A left panel shows AAV-CRISPRa-mediated transcriptional activation of Cd80, Light, CxcllO, Gitrl, 41bbl, and IFNg, normalized to vector-transduced controls; Right panel shows Co-transfection of or Dual AAV-CRISPRa-mediated transcriptional activation of Cd80, Light, CxcllO, Gitrl, 41bbl, and IFNg, normalized to control. The results showed expression of CD80, 41BBL, and Gitrl in dual-AAV-CRISPRa infected cells assessed by flow cytometry (Fig.10B). Therapeutic effects of intratumoral injected AAV-CRISPRa with dual-AAV mediated activation of Cd80, Iight/Tnfsfl4, CxcllO, 4- 1BBL/Tnfsf9, and Ifng (CLC4I) are shown in Fig. IOC. Growth curves showed the time course sizes of E0771 tumors treated with PBS, AAV-dCas9 + Vector, AAV-dCas9 + CLCG4 (dual AAV delivered CRISRPa of Clcg)(Fig. IOC). QPCR results showed successful overexpression of Cd80, Light, CxcllO, 41bbl, or/and IFNg using AAV polycistronic ORFs (Fig. 10D). Both transfection and AAV infection led to high levels of expression of these molecules. Results showed significant therapeutic effects of intratumoral injected AAV-ORFs expressing Cd80, Light, CxcllO, 41bbl (orfClc4), or AAV expressing Cd80, Light, CxcllO, 41bbl, Ifng (orfClc4I) (Fig. 10E). Significant therapeutic effects of intratumoral injected AAV-ORFs expressing Cd80, Light, CxcllO, 41bbl, Ifng (orfClc4I) were also demonstrated (Fig. 10F).

Example 8: Immune characterization of APCM treated tumors

Immune profiling was performed to characterize AAV-mediated costimulatory molecule expression that promotes tumor immune infiltration (Figs. 11 A-l IF). Results showed flow cytometry quantification of CD45+ immune cells in tumor microenvironment at DPI = 30, showing percentages of CD45+ immune cells out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I (Fig. 11 A). The percentages of CD8+ T cells out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I are shown in Fig. 1 IB. The percentages of of CD4+ T cells out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I are shown in Fig. llC. Flow cytometry quantification of PD1+CD8+ T cells out of total CD8+ T cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV-orfClc4, AAV-orfClc4I is shown in Fig. 1 ID.) Flow cytometry quantification of monocytes out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV-Clcg4, AAV- orfClc4, AAV-orfClc4I is shown in Fig. 1 IE. Flow cytometry quantification of monocytes out of total cells in tumors treated with PBS, dual-AAV-vector, dual-AAV- Clcg4, AAV-orfClc4, AAV-orfClc4I is shown in Fig. 1 IF.

Example 9: Rational design of off-the-shelf immune gene therapy by composition subtraction (“minus-one experiment”)

Composition subtraction and testing were performed to optimize the combinations of immune-stimulating molecules for tumor immune-gene therapy (Figs. 12A-12F). Results demonstrated the therapeutic effects of intra-tumoral injected AAV-CRISPR activating Cd80, light, CxcllO, 4-1BBL, Gitrl and Ifng (Clcg4I) (Fig. 12A). Growth curves of E0771 tumors treated with PBS (n = 5), AAV- Vector (n = 5), AAV-Clcg4I are shown in Figs. 12B-12C. The “minus-one” experiment further optimized the combinatorial genes pool by removing one gene in each pool. Results showed the differential therapeutic efficacy of various different constructs of the full set and the “minus-one” constructs, with growth curves of E0771 tumors treated with PBS (n = 4), AAV-Vector (n = 4), AAV-Clcg4I (n = 4), or AAV-Clcg4Iminusl (n = 4) (Fig. 12B). Growth curves of E0771 tumors treated with AAV-Clcg4I (n = 4), or AAV-Clcg4Iminusl (n = 4) are shown in Fig. 12C. Growth curves of B16F10 melanoma treated with PBS (n = 4), AAV-Vector (n = 5), AAV-p67:Clc4I (Cd80, Light, CxcllO, 41bbl, Ifng; n = 5), AAV-P72:IFNg (n = 4) are shown in Fig. 12D. Growth curves of E0771 tumors treated with PBS (n = 6), AAV-Vector (n = 6), AAV-P72:IFNg (n = 6), AAV-p94:IClG4 (Ifng, Cd80, Light, Gitrl, 41bbl; n = 6) are shown in Fig. 12E. Growth curves of E0771 tumors treated with AAV-Vector (n = 5), AAV-polycistronic PI 19:IFNg-STOP-CD80-Light- CxcllO-41BBL (n = 5), AAV-polycistronic P 118 :IFNg-CD80-Light-Cxcl 10-41BBL (n = 5), AAV-polycistronic P100:IFNg-CD80-Light-Cxcll0 (n = 5), AAV-P67: CD80-Light- CxcllO-41BBL-IFNg (n = 5) are shown in Fig. 12F.

Example 10: Analysis of expression levels of IFNg and CD80 in mammalian cells using different polvcistronic AAV-ORFs AAV-ORFs-mediated expression of IFNg was demonstrated in different AAV constructs transfected into E0771 cells (Fig. 13 A). The percentage of IFNg expressing cells in different constructs-transfected E0771 cells is shown in Fig. 13 A, lower panel left. Fig. 13 A, low panel (right) shows the median fluorescent intensity (MFI) of IFNg- APC in different constructs-transfected E0771 cells. Fig. 13B, upper panel shows representative flow cytometry plots demonstrating AAV-ORFs-mediated expression of CD80 in different AAV constructs transfected E0771 cells. Fig. 13B lower panel (left) shows the percentage of CD80+ cells in different constructs-transfected E0771 cells and Fig. 13B lower panel (right) shows the median fluorescent intensity (MFI) of CD80-PE in different constructs-transfected E0771 cells.

Example 11 : Rational design of single AAV vectors for tumor immune gene therapies The costimulatory molecule combinations in single AAV vectors as tumor immune gene therapies were optimized (Figs. 14A-14F). Fig. 14A-14B upper panels show representative flow cytometry plots demonstrating AAV-ORFs-mediated expression of CD80, 41BBL, IL23 and IFNg in AAV-infected E0771 or MA1NC cells. Figs. 14A-14B lower panels show the percentage of CD80, 41BBL, IL23, and IFNg positive cells in different AAV-infected E0771 and MA1NC cells. Result showed significant efficacy of indicated single AAV vectors with various defined combinations of transgenes expressing immune molecules as tumor immune gene therapies, where the tumor growth curves show the dynamics of E0771 tumors treated with PBS (n = 5),

AAV- Vector (n = 10), AAV-IL23 (n = 5), AAV-IFNg (n = 6), Pooled AAVs(IFNg + CD80 + LIGHT + CXCL10 + 41BBL; n = 5) (Fig. 14C). Significant efficacy of indicated single AAV vectors was also demonstrated with various defined combinations of transgenes expressing immune molecules as tumor immune gene therapies, where the tumor growth curves show the dynamics of tumor growth curves of E0771 tumors treated AAV- Vector (n =5), AAV-IFNg (n = 4), AAV-IFNg+AAV-IL23 (n = 5), pooled AAVs (IFNg + CXCL10 + 41BBL; n = 5), pooled AAVs (IFNg + LIGHT + 41BBL; n = 5), pooled AAV(IFNg + LIGHT + CXCL10 + 41BBL; n = 5) (Fig. 14D). Significant efficacy of indicated single AAV vectors was demonstrated with various defined combinations of transgenes expressing immune molecules as tumor immune gene therapies, where the tumor growth curves show the dynamics of E0771 tumors treated AAV- Vector (n =11), AAV-IFNg (n = 10), AAV-IL23 (n = 10), AAV-IFNg+AAV-IL23 (n = 4) (Fig. 14E). Tumor growth curves of E0771 tumors treated AAV-Vector (n = 5), AAV-IFNg (n = 5), AAV-IL23 (n = 5), AAV-IFNg+AAV-IL23+AAV-41BBL (n = 5) are shown in Fig. 14F.

Other Embodiments The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or subcombination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. It is to be understood that wherever values and ranges are provided herein, all values and ranges encompassed by these values and ranges, are meant to be encompassed within the scope of the present invention. Moreover, all values that fall within these ranges, as well as the upper or lower limits of a range of values, are also contemplated by the present application. The disclosures of each and every patent, patent application, and publication cited herein are hereby incorporated herein by reference in their entirety. While this invention has been disclosed with reference to specific embodiments, it is apparent that other embodiments and variations of this invention may be devised by others skilled in the art without departing from the true spirit and scope of the invention. The appended claims are intended to be construed to include all such embodiments and equivalent variations.