CA3157707A1 - Variants of cas12a nucleases and methods of making and use thereof - Google Patents

Variants of cas12a nucleases and methods of making and use thereof

Info

Publication number
CA3157707A1
CA3157707A1 CA3157707A CA3157707A CA3157707A1 CA 3157707 A1 CA3157707 A1 CA 3157707A1 CA 3157707 A CA3157707 A CA 3157707A CA 3157707 A CA3157707 A CA 3157707A CA 3157707 A1 CA3157707 A1 CA 3157707A1
Authority
CA
Canada
Prior art keywords
sequence
nucleic acid
activity
polypeptide
lbcas12a
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CA3157707A
Other languages
English (en)
French (fr)
Inventor
Joseph Matthew Watts
Sathya Sheela JALI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Pairwise Plants Services Inc
Original Assignee
Pairwise Plants Services Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Pairwise Plants Services Inc filed Critical Pairwise Plants Services Inc
Publication of CA3157707A1 publication Critical patent/CA3157707A1/en
Pending legal-status Critical Current

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/16Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/20Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Spirochaetales (O), e.g. Treponema, Leptospira
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/102Mutagenizing nucleic acids
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/10Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034Isolating an individual clone by screening libraries
    • C12N15/1093General methods of preparing gene libraries, not provided for in other subgroups
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09Recombinant DNA-technology
    • C12N15/11DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113Non-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
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14Hydrolases (3)
    • C12N9/78Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/70Fusion polypeptide containing domain for protein-protein interaction
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00Structure or type of the nucleic acid
    • C12N2310/10Type of nucleic acid
    • C12N2310/20Type of nucleic acid involving clustered regularly interspaced short palindromic repeats [CRISPR]
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y305/00Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
    • C12Y305/04Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amidines (3.5.4)
    • C12Y305/04004Adenosine deaminase (3.5.4.4)
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12YENZYMES
    • C12Y305/00Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
    • C12Y305/04Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in cyclic amidines (3.5.4)
    • C12Y305/04005Cytidine deaminase (3.5.4.5)

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (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)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Plant Pathology (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)
CA3157707A 2019-10-17 2020-10-15 Variants of cas12a nucleases and methods of making and use thereof Pending CA3157707A1 (en)

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 (1)

Publication Number Publication Date
CA3157707A1 true CA3157707A1 (en) 2021-04-22

Family

ID=75492833

Family Applications (1)

Application Number Title Priority Date Filing Date
CA3157707A Pending CA3157707A1 (en) 2019-10-17 2020-10-15 Variants of cas12a nucleases and methods of making and use thereof

Country Status (12)

Country Link
US (3) US11866745B2 (https=)
EP (1) EP4045643A4 (https=)
JP (2) JP7644755B2 (https=)
KR (1) KR20220097408A (https=)
CN (1) CN114829595A (https=)
AU (1) AU2020366358A1 (https=)
BR (1) BR112022007125A2 (https=)
CA (1) CA3157707A1 (https=)
CL (4) CL2022000937A1 (https=)
IL (1) IL292117A (https=)
MX (1) MX2022004549A (https=)
WO (1) WO2021076682A1 (https=)

Families Citing this family (29)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3204496A1 (en) 2014-10-10 2017-08-16 Editas Medicine, Inc. Compositions and methods for promoting homology directed repair
GB201506509D0 (en) 2015-04-16 2015-06-03 Univ Wageningen Nuclease-mediated genome editing
CN110382692A (zh) * 2016-04-19 2019-10-25 博德研究所 新型crispr酶以及系统
WO2018022634A1 (en) 2016-07-26 2018-02-01 The General Hospital Corporation Variants of crispr from prevotella and francisella 1 (cpf1)
WO2022174108A1 (en) * 2021-02-12 2022-08-18 The Board Of Trustees Of The Leland Stanford Junior University Synthetic cas12a for enhanced 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
CA3230609A1 (en) * 2021-09-08 2023-03-16 Shaorong Chong 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
IL314074A (en) * 2022-01-06 2024-09-01 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
AR129505A1 (es) * 2022-06-01 2024-09-04 Inst Genetics & Developmental Biology Cas Novedoso sistema de edición de genes crispr
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
CN116144631B (zh) * 2023-01-17 2023-09-15 华中农业大学 耐热型核酸内切酶及其介导的基因编辑系统
WO2024182746A1 (en) 2023-03-01 2024-09-06 Pairwise Plants Services, Inc. Engineered proteins and methods of use thereof
CN116410955B (zh) * 2023-03-10 2023-12-19 华中农业大学 两种新型核酸内切酶及其在核酸检测中的应用
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
WO2025007140A2 (en) 2023-06-30 2025-01-02 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
US20250319206A1 (en) 2024-04-04 2025-10-16 Christina Care Gene Editing Institute, Inc. Crispr/cas gene editing of neh4 and/or neh5 domains in nrf2
WO2025226816A1 (en) 2024-04-23 2025-10-30 Christiana Care Gene Editing Institute, Inc. Methods of identifying and correlating crispr-induced exon skipping to phenotypic outcomes
WO2025244732A1 (en) * 2024-05-24 2025-11-27 The Regents Of The University Of California Variant cas12a protein compositions and methods
US20260009010A1 (en) 2024-07-08 2026-01-08 Pairwise Plants Services, Inc. Methods and compositions for modification of protospacer adjacent motif specificity of cas12a
US12428636B1 (en) 2024-07-08 2025-09-30 Pairwise Plants Services, Inc. Methods and compositions for modification of protospacer adjacent motif specificity of CAS12A
CN119506250B (zh) * 2024-11-29 2025-09-09 大连理工大学 一种多PAM选择的基因编辑系统NovaCas-2及其应用
CN119876148B (zh) * 2025-01-23 2025-10-17 中国科学技术大学 Crispr rna突变体及其在基因编辑和核酸检测中的应用
CN121653102A (zh) * 2026-02-09 2026-03-13 武汉艾迪晶生物科技有限公司 一种Cas酶突变体及组合蛋白、核酸分子、重组载体、转基因细胞及其应用

Family Cites Families (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9790490B2 (en) * 2015-06-18 2017-10-17 The Broad Institute Inc. CRISPR enzymes and systems
WO2017106657A1 (en) * 2015-12-18 2017-06-22 The Broad Institute Inc. Novel crispr enzymes and systems
WO2017127807A1 (en) * 2016-01-22 2017-07-27 The Broad Institute Inc. Crystal structure of crispr cpf1
CN110382692A (zh) 2016-04-19 2019-10-25 博德研究所 新型crispr酶以及系统
CN117925730A (zh) * 2016-04-21 2024-04-26 国立大学法人神户大学 提高基因组序列修饰技术中的突变导入效率的方法、及其使用的分子复合体
CA3038960A1 (en) * 2016-09-30 2018-04-05 The Regents Of The University Of California Rna-guided nucleic acid modifying enzymes and methods of use thereof
CA3057192A1 (en) 2017-03-23 2018-09-27 President And Fellows Of Harvard College Nucleobase editors comprising nucleic acid programmable dna binding proteins
WO2018195545A2 (en) * 2017-04-21 2018-10-25 The General Hospital Corporation Variants of cpf1 (cas12a) with altered pam specificity
EP3625342B1 (en) 2017-05-18 2022-08-24 The Broad Institute, Inc. Systems, methods, and compositions for targeted nucleic acid editing
WO2018213708A1 (en) 2017-05-18 2018-11-22 The Broad Institute, Inc. Systems, methods, and compositions for targeted nucleic acid editing
JP2019016859A (ja) 2017-07-04 2019-01-31 オリンパス株式会社 観察装置および観察方法
CN109957569B (zh) * 2017-12-22 2022-10-25 苏州齐禾生科生物科技有限公司 基于cpf1蛋白的碱基编辑系统和方法
BR112020014017A2 (pt) 2018-01-11 2020-12-08 KWS SAAT SE & Co. KGaA Sistemas crispr / cpf1 de plantas otimizados
CN108486146B (zh) * 2018-03-16 2021-02-19 中国农业科学院作物科学研究所 LbCpf1-RR突变体用于CRISPR/Cpf1系统在植物基因编辑中的应用

Also Published As

Publication number Publication date
US20240076639A1 (en) 2024-03-07
WO2021076682A1 (en) 2021-04-22
CL2024001737A1 (es) 2024-11-15
US20240076640A1 (en) 2024-03-07
CN114829595A (zh) 2022-07-29
IL292117A (en) 2022-06-01
JP7644755B2 (ja) 2025-03-12
AU2020366358A1 (en) 2022-04-28
JP2025087754A (ja) 2025-06-10
CL2024001734A1 (es) 2024-11-15
US11866745B2 (en) 2024-01-09
KR20220097408A (ko) 2022-07-07
NZ787124A (en) 2025-09-26
BR112022007125A2 (pt) 2022-07-05
MX2022004549A (es) 2022-07-21
US20210115421A1 (en) 2021-04-22
EP4045643A1 (en) 2022-08-24
JP2022552409A (ja) 2022-12-15
CL2022000937A1 (es) 2023-01-20
EP4045643A4 (en) 2023-11-08
CL2024001733A1 (es) 2024-11-15

Similar Documents

Publication Publication Date Title
US20240076639A1 (en) Variants of cas12a nucleases and methods of making and use thereof
US12084671B2 (en) Optimized CRISPR-Cas nucleases and base editors and methods of use thereof
EP4471149A2 (en) Type v crispr-cas base editors and methods of use thereof
US20210130835A1 (en) Compositions and methods for rna-encoded dna-replacement of alleles
US20210238598A1 (en) Compositions, systems, and methods for base diversification
US20240327820A1 (en) Compositions, systems, and methods for base diversification
US12173335B2 (en) Recruitment of DNA polymerase for templated editing
US20260078360A1 (en) Engineered proteins and methods of use thereof
US20240327811A1 (en) Engineered proteins and methods of use thereof
US20240200102A1 (en) Fusion proteins comprising an intein polypeptide and methods of use thereof
US12428636B1 (en) Methods and compositions for modification of protospacer adjacent motif specificity of CAS12A
US20260022361A1 (en) Engineered proteins and methods of use thereof

Legal Events

Date Code Title Description
EEER Examination request

Effective date: 20220411

D15 Examination report completed

Free format text: ST27 STATUS EVENT CODE: A-2-2-D10-D15-D126 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: EXAMINER'S REPORT

Effective date: 20240808

MFA Maintenance fee for application paid

Free format text: FEE DESCRIPTION TEXT: MF (APPLICATION, 4TH ANNIV.) - STANDARD

Year of fee payment: 4

U00 Fee paid

Free format text: ST27 STATUS EVENT CODE: A-2-2-U10-U00-U101 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: MAINTENANCE REQUEST RECEIVED

Effective date: 20240930

U11 Full renewal or maintenance fee paid

Free format text: ST27 STATUS EVENT CODE: A-2-2-U10-U11-U102 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: MAINTENANCE FEE PAYMENT DETERMINED COMPLIANT

Effective date: 20240930

Free format text: ST27 STATUS EVENT CODE: A-2-2-U10-U11-U102 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: MAINTENANCE FEE PAYMENT PAID IN FULL

Effective date: 20240930

P11 Amendment of application requested

Free format text: ST27 STATUS EVENT CODE: A-2-2-P10-P11-P100 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: AMENDMENT RECEIVED - RESPONSE TO EXAMINER'S REQUISITION

Effective date: 20241105

W00 Other event occurred

Free format text: ST27 STATUS EVENT CODE: A-2-2-W10-W00-W111 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: CORRESPONDENT DETERMINED COMPLIANT

Effective date: 20250129

P11 Amendment of application requested

Free format text: ST27 STATUS EVENT CODE: A-2-2-P10-P11-P102 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: AMENDMENT DETERMINED COMPLIANT

Effective date: 20250528

P13 Application amended

Free format text: ST27 STATUS EVENT CODE: A-2-2-P10-P13-X000 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: APPLICATION AMENDED

Effective date: 20250528

D15 Examination report completed

Free format text: ST27 STATUS EVENT CODE: A-2-2-D10-D15-D126 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: EXAMINER'S REPORT

Effective date: 20251007

W00 Other event occurred

Free format text: ST27 STATUS EVENT CODE: A-2-2-W10-W00-W100 (AS PROVIDED BY THE NATIONAL OFFICE); EVENT TEXT: LETTER SENT

Effective date: 20251126