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Title:
ENGINEERED CRISPR/CAS13 SYSTEM AND USES THEREOF
Document Type and Number:
WIPO Patent Application WO/2022/068912
Kind Code:
A1
Abstract:
The invention provides novel engineered CRISPR/Cas effector enzymes, such as Cas13 (e.g., Cas13d, Cas13e, or Cas13f) that substantially maintain guide-sequence-specific endonuclease activity and substantially lack guide-sequence-independent collateral endonuclease activity compared to the corresponding wild-type Cas. Also provided are polynucleotides encoding the same, vectors or host cells comprising the polynucleotides or engineered Cas, and method of use, such as in RNA-based target gene transcript knock down.

Inventors:
TONG HUAWEI (CN)
WANG XING (CN)
WANG SHAORAN (CN)
Application Number:
PCT/CN2021/121926
Publication Date:
April 07, 2022
Filing Date:
September 29, 2021
Export Citation:
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Assignee:
HUIGENE THERAPEUTICS CO LTD (CN)
International Classes:
C12N15/11; C12N9/22
Domestic Patent References:
WO2020028555A22020-02-06
WO2019040664A12019-02-28
WO2017219027A12017-12-21
WO2020028555A22020-02-06
WO2017219027A12017-12-21
WO2016205764A12016-12-22
WO2014093622A22014-06-19
WO2015070083A12015-05-14
WO2016094872A12016-06-16
WO2017070605A12017-04-27
WO2016205764A12016-12-22
Foreign References:
CN112410377A2021-02-26
US8871445B22014-10-28
US20160208243A12016-07-21
US20130074667W2013-12-12
US9790490B22017-10-17
US8795965B22014-08-05
US8795965B22014-08-05
JPS501B11975-01-06
US20170362644A12017-12-21
EP3009511A22016-04-20
US8454972B22013-06-04
US5593972A1997-01-14
US5589466A1996-12-31
US5580859A1996-12-03
Other References:
EAST-SELETSKY ALEXANDRA ET AL: "RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes", MOLECULAR CELL, ELSEVIER, AMSTERDAM , NL, vol. 66, no. 3, 4 May 2017 (2017-05-04), pages 373, XP029999996, ISSN: 1097-2765, DOI: 10.1016/J.MOLCEL.2017.04.008
MICHAL BURMISTRZ ET AL: "RNA-Targeting CRISPR-Cas Systems and Their Applications", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 21, no. 3, 7 February 2020 (2020-02-07), pages 1122, XP055726418, DOI: 10.3390/ijms21031122
OMAR O ABUDAYYEH ET AL: "RNA targeting with CRISPR-Cas13", NATURE, vol. 550, no. 7675, 4 October 2017 (2017-10-04), London, pages 280 - 284, XP055529736, ISSN: 0028-0836, DOI: 10.1038/nature24049
DAVID B. T. COX ET AL: "RNA editing with CRISPR-Cas13", SCIENCE, vol. 358, no. 6366, 24 November 2017 (2017-11-24), US, pages 1019 - 1027, XP055491658, ISSN: 0036-8075, DOI: 10.1126/science.aaq0180
HE ET AL., PROTEIN CELL, vol. 11, 2020, pages 518 - 524
ZHOU ET AL., CELL, vol. 181, 2020, pages 590 - 603
ZHOU ET AL., , NATIONAL SCIENCE, vol. 7, 2020, pages 835 - 837
WRIGHT E: "Rational design of a split-Cas9 enzyme complex", PROC. NAT'L. ACAD. SCI., vol. 112, no. 10, 2015, pages 2984 - 2989, XP055283739, DOI: 10.1073/pnas.1501698112
HIROSAWA ET AL., NUCL. ACIDS RES., vol. 45, no. 13, 2017, pages 118
KONERMANN ET AL.: "Optical control of mammalian endogenous transcription and epigenetic states", NATURE, vol. 500, 2013, pages 7463
ZETSCHE ET AL.: "A split-Cas9 architecture for inducible genome editing and transcription modulation", NATURE BIOTECH, vol. 33, no. 2, 2015, pages 139 - 42, XP055227889, DOI: 10.1038/nbt.3149
GOLDFLESS ET AL.: "Direct and specific chemical control of eukaryotic translation with a synthetic RNA-protein interaction", NUCL. ACIDS RES., vol. 40, no. 9, 2012, pages e64 - e64, XP055433199, DOI: 10.1093/nar/gks028
NAKAMURA ET AL., NUCL. ACIDS RES., vol. 28, 2000, pages 292
NAKAMURA, Y. ET AL.: "Codon usage tabulated from the international DNA sequence databases: status for the year 2000", NUCL. ACIDS RES, vol. 28, 2000, pages 292, XP002941557, DOI: 10.1093/nar/28.1.292
GRISSA ET AL., NUCLEIC ACIDS RES., vol. 35, 2007, pages W52 - 7
GRISSA ET AL., BMC BIOINFORMATICS, vol. 8, 2007, pages 172
GRISSA ET AL., NUCLEIC ACIDS RES., vol. 36, 2008, pages W145 - 8
MOLLERLIANG, PEERJ, vol. 5, 2017, pages e3788
ECKSTEIN: "Phosphorothioates, essential components of therapeutic oligonucleotides", NUCL. ACID THER.,, vol. 24, 2014, pages 374 - 387, XP055518661, DOI: 10.1089/nat.2014.0506
ALLERSON ET AL.: "Fully 2'-modified oligonucleotide duplexes with improved in vitro potency and stability compared to unmodified small interfering RNA", J. MED. CHEM., vol. 48, no. 4, 2005, pages 901 - 904, XP055674720, DOI: 10.1021/jm049167j
BRAMSEN ET AL.: "Development of therapeutic-grade small interfering RNAs by chemical engineering", FRONT. GENET., vol. 20, no. 3, August 2012 (2012-08-01), pages 154
KELLEY ET AL.: "Versatility of chemically synthesized guide RNAs for CRISPR-Cas9 genome editing", J. BIOTECHNOL., vol. 233, 2016, pages 74 - 83, XP029674969, DOI: 10.1016/j.jbiotec.2016.06.011
NOWAK ET AL.: "Guide RNA engineering for versatile Cas9 functionality", NUCL. ACID. RES., vol. 44, no. 20, 2016, pages 9555 - 9564, XP055524584, DOI: 10.1093/nar/gkw908
HENDEL, NAT BIOTECHNOL., vol. 33, no. 9, 2015, pages 985 - 9
GOOTENBERG ET AL.: "Nucleic acid detection with CRISPR-Casl3a/C2c2", SCIENCE, vol. 356, no. 6336, 28 April 2017 (2017-04-28), pages 438 - 442, XP055781069, DOI: 10.1126/science.aam9321
CHEN ET AL., SCIENCE, vol. 348, no. 6233, 24 April 2015 (2015-04-24), pages 6090
BELL ET AL.: "A high-throughput screening strategy for detecting CRISPR-Cas9 induced mutations using next-generation sequencing", BMC GENOMICS, vol. 15, no. 1, 2014, pages 1002, XP021203078, DOI: 10.1186/1471-2164-15-1002
VERWAAL ET AL.: "CRISPR/Cpfl enables fast and simple genome editing of Saccharomyces cerevisiae", YEAST, 2017
HLAVOVA ET AL.: "Improving microalgae for biotechnology-from genetics to synthetic biology", BIOTECHNOL. ADV, vol. 33, 2015, pages 1194 - 203
NICOLAOU ET AL.: "Molecular diagnosis of peanut and legume allergy", CURR. OPIN. ALLERGY CLIN. IMMUNOL., vol. 11, no. 3, 2011, pages 222 - 8
DATLINGER ET AL.: "Pooled CRISPR screening with single-cell transcriptome read-out", NAT. METHODS., vol. 14, no. 3, 2017, pages 297 - 301
CANVER ET AL.: "BCL11A enhancer dissection by Cas9-mediated in situ saturating mutagenesis", NATURE, vol. 527, no. 7577, 2015, pages 192 - 7, XP055274680, DOI: 10.1038/nature15521
OSBORNE ET AL.: "RNA-dominant diseases", HUM. MOL. GENET., vol. 18, no. 8, 15 April 2009 (2009-04-15), pages 1471 - 81
COOPER ET AL.: "RNA and disease", CELL, vol. 136, no. 4, 2009, pages 777 - 793
HALLBRINK ET AL.: "Prediction of cell-penetrating peptides", METHODS MOL. BIOL.,, vol. 1324, 2015, pages 39 - 58
RAMAKRISHNA ET AL.: "Gene disruption by cell-penetrating peptide-mediated delivery of Cas9 protein and guide RNA", GENOME RES., vol. 24, no. 6, June 2014 (2014-06-01), pages 1020 - 7, XP055692365, DOI: 10.1101/gr.171264.113
Attorney, Agent or Firm:
KING & WOOD MALLESONS (CN)
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