CN114829595B - Cas12a核酸酶的变体及其制备方法和用途 - Google Patents

Cas12a核酸酶的变体及其制备方法和用途

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Publication number
CN114829595B
CN114829595B CN202080087121.8A CN202080087121A CN114829595B CN 114829595 B CN114829595 B CN 114829595B CN 202080087121 A CN202080087121 A CN 202080087121A CN 114829595 B CN114829595 B CN 114829595B
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CN114829595A (zh
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J·M·瓦特斯
S·S·贾里
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Pairing Plant Service Co ltd
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    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/20—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Spirochaetales (O), e.g. Treponema, Leptospira
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102—Mutagenizing nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1093—General methods of preparing gene libraries, not provided for in other subgroups
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/20—Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
    • C12Y305/04—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amidines (3.5.4)
    • C12Y305/04004—Adenosine deaminase (3.5.4.4)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
    • C12Y305/04—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amidines (3.5.4)
    • C12Y305/04005—Cytidine deaminase (3.5.4.5)

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
CN202080087121.8A 2019-10-17 2020-10-15 Cas12a核酸酶的变体及其制备方法和用途 Active CN114829595B (zh)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201962916392P 2019-10-17 2019-10-17
US62/916,392 2019-10-17
PCT/US2020/055659 WO2021076682A1 (en) 2019-10-17 2020-10-15 Variants of cas12a nucleases and methods of making and use thereof

Publications (2)

Publication Number Publication Date
CN114829595A CN114829595A (zh) 2022-07-29
CN114829595B true CN114829595B (zh) 2026-05-29

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US (3) US11866745B2 (enExample)
EP (1) EP4045643A4 (enExample)
JP (2) JP7644755B2 (enExample)
KR (1) KR20220097408A (enExample)
CN (1) CN114829595B (enExample)
AU (1) AU2020366358A1 (enExample)
BR (1) BR112022007125A2 (enExample)
CA (1) CA3157707A1 (enExample)
CL (4) CL2022000937A1 (enExample)
IL (1) IL292117A (enExample)
MX (1) MX2022004549A (enExample)
WO (1) WO2021076682A1 (enExample)

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AU2017302551B2 (en) 2016-07-26 2023-04-27 The General Hospital Corporation Variants of CRISPR from Prevotella and Francisella 1 (Cpf1)
EP4291644A4 (en) * 2021-02-12 2026-02-18 Univ Leland Stanford Junior CAS12A SYNTHETIC FOR IMPROVED MULTIPLEX GENE CONTROL AND EDITING
CN113136376B (zh) * 2021-05-26 2022-10-21 武汉大学 一种Cas12a变体及其在基因编辑中的应用
EP4399310A4 (en) * 2021-09-08 2025-07-16 Arbor Biotechnologies Inc COMPOSITIONS COMPRISING A CRISPR NUCLEASE AND USES THEREOF
CN114214348A (zh) * 2021-11-16 2022-03-22 河南农业大学 应用于核酸检测的Crispr/LbCas12a突变体蛋白及其制备方法和应用
CN118339285A (zh) * 2021-12-09 2024-07-12 北京干细胞与再生医学研究院 工程化Cas12b效应子蛋白及其使用方法
CA3245296A1 (en) * 2022-01-06 2023-07-13 Ensoma Aps CAS12A ENDONUCLEASE VARIANTS AND METHODS OF USE
EP4508205A1 (en) * 2022-04-12 2025-02-19 John Innes Centre Compositions and methods for increasing genome editing efficiency
WO2023232109A1 (zh) * 2022-06-01 2023-12-07 中国科学院遗传与发育生物学研究所 新的crispr基因编辑系统
CN116286734B (zh) * 2022-11-29 2024-04-02 武汉大学 野生型LbCas12a蛋白的突变体及SNP检测用途
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CN116179512B (zh) * 2023-03-16 2023-09-15 华中农业大学 靶标识别范围广的核酸内切酶及其应用
AU2024309386A1 (en) 2023-06-30 2025-12-18 Christiana Care Gene Editing Institute, Inc. Nras gene knockout for treatment of cancer
KR102789465B1 (ko) 2023-09-20 2025-04-04 주식회사 엔세이지 신규 CRISPR/Cas12a 시스템 및 그 유전자 편집 용도
CN119432813A (zh) * 2023-12-29 2025-02-14 武汉艾迪晶生物科技有限公司 Cas蛋白、组合蛋白、核酸分子、核糖核蛋白复合物、重组载体、转基因细胞及其应用
CN117844782B (zh) * 2024-03-06 2024-05-31 崖州湾国家实验室 靶向范围广的基因编辑核酸酶及其在核酸检测中的应用
WO2025213016A2 (en) 2024-04-04 2025-10-09 Christiana Care Gene Editing Institute, Inc. Crispr/cas gene editing of neh4 and/or neh5 domains in nrf2
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US11866745B2 (en) 2024-01-09
KR20220097408A (ko) 2022-07-07
JP2025087754A (ja) 2025-06-10
WO2021076682A1 (en) 2021-04-22
US20210115421A1 (en) 2021-04-22
JP2022552409A (ja) 2022-12-15
CL2024001733A1 (es) 2024-11-15
US20240076639A1 (en) 2024-03-07
MX2022004549A (es) 2022-07-21
EP4045643A4 (en) 2023-11-08
CL2022000937A1 (es) 2023-01-20
US20240076640A1 (en) 2024-03-07
BR112022007125A2 (pt) 2022-07-05
CN114829595A (zh) 2022-07-29
CL2024001734A1 (es) 2024-11-15
AU2020366358A1 (en) 2022-04-28
CL2024001737A1 (es) 2024-11-15
JP7644755B2 (ja) 2025-03-12
CA3157707A1 (en) 2021-04-22
IL292117A (en) 2022-06-01
NZ787124A (en) 2025-09-26
EP4045643A1 (en) 2022-08-24

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