AU2018320865B2 - Engineered CRISPR-Cas9 nucleases with altered PAM specificity - Google Patents

Engineered CRISPR-Cas9 nucleases with altered PAM specificity Download PDF

Info

Publication number
AU2018320865B2
AU2018320865B2 AU2018320865A AU2018320865A AU2018320865B2 AU 2018320865 B2 AU2018320865 B2 AU 2018320865B2 AU 2018320865 A AU2018320865 A AU 2018320865A AU 2018320865 A AU2018320865 A AU 2018320865A AU 2018320865 B2 AU2018320865 B2 AU 2018320865B2
Authority
AU
Australia
Prior art keywords
protein
spcas9
fusion protein
pam
variants
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.)
Active
Application number
AU2018320865A
Other languages
English (en)
Other versions
AU2018320865A1 (en
Inventor
Keith J. Joung
Benjamin KLEINSTIVER
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.)
General Hospital Corp
Original Assignee
General Hospital Corp
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 General Hospital Corp filed Critical General Hospital Corp
Publication of AU2018320865A1 publication Critical patent/AU2018320865A1/en
Application granted granted Critical
Publication of AU2018320865B2 publication Critical patent/AU2018320865B2/en
Priority to AU2023278074A priority Critical patent/AU2023278074B2/en
Priority to AU2025275152A priority patent/AU2025275152B2/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/22—Ribonucleases [RNase]; Deoxyribonucleases [DNase]
    • 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
    • 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
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90—Stable introduction of foreign DNA into chromosome
    • C12N15/902—Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907—Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
    • 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
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0071—Oxidoreductases (1.) acting on paired donors with incorporation of molecular oxygen (1.14)
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1003—Transferases (2.) transferring one-carbon groups (2.1)
    • C12N9/1007—Methyltransferases (general) (2.1.1.)
    • 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
    • C12N9/10—Transferases (2.)
    • C12N9/1025—Acyltransferases (2.3)
    • C12N9/1029—Acyltransferases (2.3) transferring groups other than amino-acyl groups (2.3.1)
    • 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
    • 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)
    • C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/71—Fusion polypeptide containing domain for protein-protein interaction containing domain for transcriptional activaation, e.g. VP16
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/80—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor
    • 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
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/80—Vectors containing sites for inducing double-stranded breaks, e.g. meganuclease restriction sites

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)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biophysics (AREA)
  • Plant Pathology (AREA)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
AU2018320865A 2017-08-23 2018-08-22 Engineered CRISPR-Cas9 nucleases with altered PAM specificity Active AU2018320865B2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU2023278074A AU2023278074B2 (en) 2017-08-23 2023-12-07 Engineered CRISPR-Cas9 nucleases with altered PAM specificity
AU2025275152A AU2025275152B2 (en) 2017-08-23 2025-12-03 Engineered CRISPR-Cas9 nucleases with altered PAM specificity

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
US201762549303P 2017-08-23 2017-08-23
US62/549,303 2017-08-23
US201862641687P 2018-03-12 2018-03-12
US62/641,687 2018-03-12
PCT/US2018/047577 WO2019040650A1 (en) 2017-08-23 2018-08-22 GENETICALLY MODIFIED CRISPR-CAS9 NUCLEASES HAVING MODIFIED PAM SPECIFICITY

Related Child Applications (1)

Application Number Title Priority Date Filing Date
AU2023278074A Division AU2023278074B2 (en) 2017-08-23 2023-12-07 Engineered CRISPR-Cas9 nucleases with altered PAM specificity

Publications (2)

Publication Number Publication Date
AU2018320865A1 AU2018320865A1 (en) 2020-03-05
AU2018320865B2 true AU2018320865B2 (en) 2023-09-14

Family

ID=65439890

Family Applications (3)

Application Number Title Priority Date Filing Date
AU2018320865A Active AU2018320865B2 (en) 2017-08-23 2018-08-22 Engineered CRISPR-Cas9 nucleases with altered PAM specificity
AU2023278074A Active AU2023278074B2 (en) 2017-08-23 2023-12-07 Engineered CRISPR-Cas9 nucleases with altered PAM specificity
AU2025275152A Active AU2025275152B2 (en) 2017-08-23 2025-12-03 Engineered CRISPR-Cas9 nucleases with altered PAM specificity

Family Applications After (2)

Application Number Title Priority Date Filing Date
AU2023278074A Active AU2023278074B2 (en) 2017-08-23 2023-12-07 Engineered CRISPR-Cas9 nucleases with altered PAM specificity
AU2025275152A Active AU2025275152B2 (en) 2017-08-23 2025-12-03 Engineered CRISPR-Cas9 nucleases with altered PAM specificity

Country Status (8)

Country Link
US (3) US11286468B2 (enExample)
EP (1) EP3672612A4 (enExample)
JP (2) JP7581046B2 (enExample)
CN (2) CN111278450B (enExample)
AU (3) AU2018320865B2 (enExample)
BR (1) BR112020003596A2 (enExample)
CA (1) CA3073448A1 (enExample)
WO (1) WO2019040650A1 (enExample)

Families Citing this family (44)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9890364B2 (en) 2012-05-29 2018-02-13 The General Hospital Corporation TAL-Tet1 fusion proteins and methods of use thereof
EP3919621A1 (en) 2014-06-23 2021-12-08 The General Hospital Corporation Genomewide unbiased identification of dsbs evaluated by sequencing (guide-seq)
AU2015330699B2 (en) 2014-10-10 2021-12-02 Editas Medicine, Inc. Compositions and methods for promoting homology directed repair
JP6817215B2 (ja) 2015-03-03 2021-01-20 ザ ジェネラル ホスピタル コーポレイション 変更ＰＡＭ特異性を有する遺伝子操作ＣＲＩＳＰＲ−Ｃａｓ９ヌクレアーゼ
CN108350453A (zh) 2015-09-11 2018-07-31 通用医疗公司 核酸酶dsb的完全查询和测序(find-seq)
US11168313B2 (en) 2016-07-26 2021-11-09 The General Hospital Corporation Variants of CRISPR from Prevotella and Francisella 1 (Cpf1)
WO2018119010A1 (en) 2016-12-19 2018-06-28 Editas Medicine, Inc. Assessing nuclease cleavage
EP3565907B1 (en) 2017-01-06 2022-05-04 Editas Medicine, Inc. Methods of assessing nuclease cleavage
ES2992912T3 (en) 2017-04-21 2024-12-19 Massachusetts Gen Hospital Variants of cpf1 (cas12a) with altered pam specificity
US11499151B2 (en) 2017-04-28 2022-11-15 Editas Medicine, Inc. Methods and systems for analyzing guide RNA molecules
AU2018273986A1 (en) 2017-05-25 2019-12-12 The General Hospital Corporation Bipartite base editor (BBE) architectures and type-II-C-Cas9 zinc finger editing
JP7518620B2 (ja) 2017-06-09 2024-07-18 エディタス・メディシン，インコーポレイテッド 操作されたｃａｓ９ヌクレアーゼ
WO2019014564A1 (en) 2017-07-14 2019-01-17 Editas Medicine, Inc. SYSTEMS AND METHODS OF TARGETED INTEGRATION AND GENOME EDITING AND DETECTION THEREOF WITH INTEGRATED PRIMING SITES
CA3073448A1 (en) 2017-08-23 2019-02-28 The General Hospital Corporation Engineered crispr-cas9 nucleases with altered pam specificity
EP3781585A4 (en) 2018-04-17 2022-01-26 The General Hospital Corporation SENSITIVE IN VITRO TESTS FOR SUBSTRATE PREFERENCES AND SITE FOR NUCLEIC ACID BINDERS, MODIFIERS AND CLEAVAGE AGENTS
AU2019265019B2 (en) 2018-05-11 2025-11-06 Beam Therapeutics Inc. Methods of substituting pathogenic amino acids using programmable base editor systems
KR20250133482A (ko) * 2018-05-11 2025-09-05 빔 테라퓨틱스, 인크. 프로그래밍가능한 염기 편집기 시스템을 이용하여 단일 뉴클레오티드 다형성을 편집하는 방법
AU2019291918B2 (en) 2018-06-29 2025-06-12 Editas Medicine, Inc. Synthetic guide molecules, compositions and methods relating thereto
AU2019336245B2 (en) 2018-09-07 2026-02-26 Beam Therapeutics Inc. Compositions and methods for delivering a nucleobase editing system
EP3850088A4 (en) 2018-09-07 2023-07-19 Beam Therapeutics, Inc. Compositions and methods for improving base editing
CN114040977B (zh) 2019-01-31 2024-09-03 比姆医疗股份有限公司 非靶脱氨反应减低的核碱基编辑器和用于定性核碱基编辑器的测定
EP3921417A4 (en) 2019-02-04 2022-11-09 The General Hospital Corporation VARIANTS OF ADENINE DNA BASES WITH REDUCED OFF-TARGET RNA EDIT
AU2020223314A1 (en) 2019-02-13 2021-08-12 Beam Therapeutics Inc. Compositions and methods for treating alpha-1 antitrypsin deficiency
SG11202108346WA (en) 2019-02-13 2021-08-30 Beam Therapeutics Inc Modified immune cells having adenosine deaminase base editors for modifying a nucleobase in a target sequence
CN112266418A (zh) * 2019-07-08 2021-01-26 中国科学院微生物研究所 改进的基因组编辑系统及其应用
CN112280771A (zh) * 2019-07-10 2021-01-29 中国科学院遗传与发育生物学研究所 双功能基因组编辑系统及其用途
EP4021945A4 (en) * 2019-08-30 2023-11-15 The General Hospital Corporation Combinatorial adenine and cytosine dna base editors
AU2020345830A1 (en) * 2019-09-09 2022-03-24 Beam Therapeutics Inc. Novel CRISPR enzymes, methods, systems and uses thereof
CN115279398B (zh) 2019-09-27 2025-07-04 比姆医疗股份有限公司 治疗液态癌症的组合物和方法
US12497609B2 (en) 2019-10-14 2025-12-16 Yonsei University Biohealth Technology Holdings. Inc. Protospacer adjacent motif sequence and method for modifying target nucleic acid in genome of cell by using same
WO2021151085A2 (en) 2020-01-24 2021-07-29 The General Hospital Corporation Crispr-cas enzymes with enhanced on-target activity
WO2021151073A2 (en) * 2020-01-24 2021-07-29 The General Hospital Corporation Unconstrained genome targeting with near-pamless engineered crispr-cas9 variants
EP4114937A4 (en) 2020-03-04 2024-09-04 Flagship Pioneering Innovations VI, LLC METHODS AND COMPOSITIONS FOR MODULATING A GENOME
US12576151B2 (en) 2020-09-25 2026-03-17 Beam Therapeutics Inc. Fratricide resistant modified immune cells and methods of using the same
CN112430613A (zh) * 2020-12-08 2021-03-02 安徽省农业科学院水稻研究所 一种宽编辑范围的SpG基因及其应用
WO2022152746A1 (en) 2021-01-13 2022-07-21 Alia Therapeutics Srl K526d cas9 variants and applications thereof
KR20260037664A (ko) 2021-09-08 2026-03-18 플래그쉽 파이어니어링 이노베이션스 브이아이, 엘엘씨 게놈을 조절하기 위한 방법 및 조성물
WO2023039435A2 (en) 2021-09-08 2023-03-16 Flagship Pioneering Innovations Vi, Llc Pah-modulating compositions and methods
CN116286736B (zh) * 2022-01-17 2026-05-01 四川大学 一种具有扩大的pam识别范围的融合蛋白及其应用
WO2025087413A1 (zh) * 2023-10-25 2025-05-01 上海津曼特生物科技有限公司 靶向ttr的基因编辑组合物
WO2025158391A2 (en) * 2024-01-24 2025-07-31 The General Hospital Corporation Generation of custom crispr-cas9 pam variant enzymes
CN118048343B (zh) * 2024-03-01 2025-01-28 上海交通大学 Cas9核酸酶及其制备方法和用途
CN120005854B (zh) * 2025-04-18 2025-08-05 广州瑞风生物科技有限公司 Cas9突变体、基因编辑系统及应用
CN120665840A (zh) * 2025-05-30 2025-09-19 浙江大学 Cas9蛋白突变体及其应用

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016141224A1 (en) * 2015-03-03 2016-09-09 The General Hospital Corporation Engineered crispr-cas9 nucleases with altered pam specificity
WO2017070633A2 (en) * 2015-10-23 2017-04-27 President And Fellows Of Harvard College Evolved cas9 proteins for gene editing

Family Cites Families (104)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005090607A1 (en) 2004-03-08 2005-09-29 Rubicon Genomics, Inc. Methods and compositions for generating and amplifying dna libraries for sensitive detection and analysis of dna methylation
CA2615323A1 (en) 2005-06-06 2007-12-21 454 Life Sciences Corporation Paired end sequencing
ATE510930T1 (de) 2005-08-02 2011-06-15 Rubicon Genomics Inc Zusammensetzungen und verfahren zur bearbeitung und amplifikation von dna mit verwendung mehrerer enzyme in einer einzigen reaktion
NZ597299A (en) 2005-08-26 2013-03-28 Dupont Nutrition Biosci Aps Use of a cas gene in combination with CRISPR repeats for modulating resistance in a cell
WO2007059041A2 (en) 2005-11-11 2007-05-24 Pfizer, Inc. Combinations and methods of using an immunomodulatory oligodeoxynucleotide
PT2126130E (pt) 2007-03-02 2015-08-03 Dupont Nutrition Biosci Aps Culturas com resistência melhorada a fagos
WO2009027978A1 (en) 2007-08-30 2009-03-05 Hadasit Medical Research Services & Development Ltd. NUCLEIC ACID SEQUENCES COMPRISING NF-ϰB BINDING SITE WITHIN O(6)-METHYLGUANINE-DNA-METHYLTRANSFERASE (MGMT) PROMOTER REGION AND USES THEREOF FOR THE TREATMENT OF CANCER AND IMMUNE-RELATED DISORDERS
US8029993B2 (en) 2008-04-30 2011-10-04 Population Genetics Technologies Ltd. Asymmetric adapter library construction
WO2010011961A2 (en) 2008-07-25 2010-01-28 University Of Georgia Research Foundation, Inc. Prokaryotic rnai-like system and methods of use
US8383345B2 (en) 2008-09-12 2013-02-26 University Of Washington Sequence tag directed subassembly of short sequencing reads into long sequencing reads
US20100076057A1 (en) 2008-09-23 2010-03-25 Northwestern University TARGET DNA INTERFERENCE WITH crRNA
US9404098B2 (en) 2008-11-06 2016-08-02 University Of Georgia Research Foundation, Inc. Method for cleaving a target RNA using a Cas6 polypeptide
EP2367938B1 (en) 2008-12-12 2014-06-11 DuPont Nutrition Biosciences ApS Genetic cluster of strains of streptococcus thermophilus having unique rheological properties for dairy fermentation
JP5387839B2 (ja) 2009-09-04 2014-01-15 アイシン・エィ・ダブリュ株式会社 ナビゲーション装置、ナビゲーション方法、及びナビゲーションプログラム
US9023769B2 (en) 2009-11-30 2015-05-05 Complete Genomics, Inc. cDNA library for nucleic acid sequencing
US9493827B2 (en) 2010-01-14 2016-11-15 Deutsches Krebsforschungszentrum Determination of in vivo DNA double-strand break localization and application thereof
JP2013518602A (ja) 2010-02-09 2013-05-23 サンガモ バイオサイエンシーズ， インコーポレイテッド 部分的に一本鎖のドナー分子による標的化ゲノム改変
US10087431B2 (en) 2010-03-10 2018-10-02 The Regents Of The University Of California Methods of generating nucleic acid fragments
EA024121B9 (ru) 2010-05-10 2017-01-30 Дзе Реджентс Ов Дзе Юниверсити Ов Калифорния Композиции эндорибонуклеаз и способы их использования
AU2011325956B2 (en) 2010-11-12 2016-07-14 The General Hospital Corporation Polycomb-associated non-coding RNAs
WO2012164565A1 (en) 2011-06-01 2012-12-06 Yeda Research And Development Co. Ltd. Compositions and methods for downregulating prokaryotic genes
WO2013066438A2 (en) 2011-07-22 2013-05-10 President And Fellows Of Harvard College Evaluation and improvement of nuclease cleavage specificity
WO2013078470A2 (en) 2011-11-22 2013-05-30 MOTIF, Active Multiplex isolation of protein-associated nucleic acids
GB201122458D0 (en) 2011-12-30 2012-02-08 Univ Wageningen Modified cascade ribonucleoproteins and uses thereof
US9637739B2 (en) 2012-03-20 2017-05-02 Vilnius University RNA-directed DNA cleavage by the Cas9-crRNA complex
EP2847353B1 (en) 2012-05-10 2022-01-19 The General Hospital Corporation Methods for determining a nucleotide sequence
LT3401400T (lt) 2012-05-25 2019-06-10 The Regents Of The University Of California Būdai ir kompozicijos, skirtos rnr molekulės nukreipiamai tikslinės dnr modifikacijai ir rnr molekulės nukreipiamam transkripcijos moduliavimui
JP6181751B2 (ja) 2012-06-18 2017-08-16 ニューゲン テクノロジーズ， インコーポレイテッド 望まれない核酸配列のネガティブ選択のための組成物および方法
US20140024542A1 (en) 2012-07-17 2014-01-23 Counsyl, Inc. Methods and compositions for enrichment of target polynucleotides
AU2012385961B9 (en) 2012-07-24 2017-05-18 Natera, Inc. Highly multiplex PCR methods and compositions
WO2014071070A1 (en) 2012-11-01 2014-05-08 Pacific Biosciences Of California, Inc. Compositions and methods for selection of nucleic acids
CN104903466B (zh) 2012-11-05 2016-11-23 鲁比康基因组学公司 条形编码核酸
EP2931898B1 (en) 2012-12-12 2016-03-09 The Broad Institute, Inc. Engineering and optimization of systems, methods and compositions for sequence manipulation with functional domains
EP2896697B1 (en) 2012-12-12 2015-09-02 The Broad Institute, Inc. Engineering of systems, methods and optimized guide compositions for sequence manipulation
JP2016504026A (ja) 2012-12-12 2016-02-12 ザ・ブロード・インスティテュート・インコーポレイテッド 配列操作のための系、方法および最適化ガイド組成物のエンジニアリング
US20140310830A1 (en) 2012-12-12 2014-10-16 Feng Zhang CRISPR-Cas Nickase Systems, Methods And Compositions For Sequence Manipulation in Eukaryotes
US8697359B1 (en) 2012-12-12 2014-04-15 The Broad Institute, Inc. CRISPR-Cas systems and methods for altering expression of gene products
WO2014093701A1 (en) 2012-12-12 2014-06-19 The Broad Institute, Inc. Functional genomics using crispr-cas systems, compositions, methods, knock out libraries and applications thereof
IL239317B (en) 2012-12-12 2022-07-01 Broad Inst Inc Providing, engineering and optimizing systems, methods and compositions for sequence manipulation and therapeutic applications
ES2576126T3 (es) 2012-12-12 2016-07-05 The Broad Institute, Inc. Modificación por tecnología genética y optimización de sistemas, métodos y composiciones enzimáticas mejorados para la manipulación de secuencias
CN114634950A (zh) 2012-12-12 2022-06-17 布罗德研究所有限公司 用于序列操纵的crispr-cas组分系统、方法以及组合物
AU2013359146B2 (en) 2012-12-13 2017-12-07 Corteva Agriscience Llc DNA detection methods for site specific nuclease activity
CA2897932A1 (en) 2013-01-14 2014-07-17 Recombinetics, Inc. Hornless livestock
US20140212869A1 (en) 2013-01-25 2014-07-31 Agilent Technologies, Inc. Nucleic Acid Proximity Assay Involving the Formation of a Three-way junction
CA2900338C (en) 2013-02-07 2024-11-05 The General Hospital Corporation TRANSCRIPTIONAL ACTIVATORS TALE
EP2958996B1 (en) 2013-02-25 2019-10-16 Sangamo Therapeutics, Inc. Methods and compositions for enhancing nuclease-mediated gene disruption
SG11201507378UA (en) 2013-03-14 2015-10-29 Caribou Biosciences Inc Compositions and methods of nucleic acid-targeting nucleic acids
US20140273230A1 (en) 2013-03-15 2014-09-18 Sigma-Aldrich Co., Llc Crispr-based genome modification and regulation
US9234213B2 (en) 2013-03-15 2016-01-12 System Biosciences, Llc Compositions and methods directed to CRISPR/Cas genomic engineering systems
US11332719B2 (en) 2013-03-15 2022-05-17 The Broad Institute, Inc. Recombinant virus and preparations thereof
US10760064B2 (en) 2013-03-15 2020-09-01 The General Hospital Corporation RNA-guided targeting of genetic and epigenomic regulatory proteins to specific genomic loci
US20140349400A1 (en) 2013-03-15 2014-11-27 Massachusetts Institute Of Technology Programmable Modification of DNA
US10378027B2 (en) 2013-03-15 2019-08-13 The General Hospital Corporation RNA-guided targeting of genetic and epigenomic regulatory proteins to specific genomic loci
RU2723130C2 (ru) 2013-04-05 2020-06-08 ДАУ АГРОСАЙЕНСИЗ ЭлЭлСи Способы и композиции для встраивания экзогенной последовательности в геном растений
DK2986729T3 (en) 2013-04-16 2018-10-29 Regeneron Pharma TARGETED MODIFICATION OF ROOT THROUGH
CA2910427C (en) 2013-05-10 2024-02-20 Sangamo Biosciences, Inc. Delivery methods and compositions for nuclease-mediated genome engineering
EP3004339B1 (en) 2013-05-29 2021-07-07 Cellectis New compact scaffold of cas9 in the type ii crispr system
WO2014204724A1 (en) 2013-06-17 2014-12-24 The Broad Institute Inc. Delivery, engineering and optimization of tandem guide systems, methods and compositions for sequence manipulation
JP6665088B2 (ja) 2013-06-17 2020-03-13 ザ・ブロード・インスティテュート・インコーポレイテッド 配列操作のための最適化されたＣＲＩＳＰＲ−Ｃａｓ二重ニッカーゼ系、方法および組成物
JP6482546B2 (ja) 2013-07-19 2019-03-13 ラリクス・バイオサイエンス・リミテッド・ライアビリティ・カンパニーＬａｒｉｘ Ｂｉｏｓｃｉｅｎｃｅ， Ｌｌｃ 二重対立遺伝子ノックアウトを生成するための方法および組成物
US20150044192A1 (en) 2013-08-09 2015-02-12 President And Fellows Of Harvard College Methods for identifying a target site of a cas9 nuclease
US9388430B2 (en) 2013-09-06 2016-07-12 President And Fellows Of Harvard College Cas9-recombinase fusion proteins and uses thereof
WO2015074017A1 (en) 2013-11-18 2015-05-21 Rubicon Genomics Degradable adaptors for background reduction
WO2015089486A2 (en) 2013-12-12 2015-06-18 The Broad Institute Inc. Systems, methods and compositions for sequence manipulation with optimized functional crispr-cas systems
WO2015089364A1 (en) 2013-12-12 2015-06-18 The Broad Institute Inc. Crystal structure of a crispr-cas system, and uses thereof
WO2015089473A1 (en) 2013-12-12 2015-06-18 The Broad Institute Inc. Engineering of systems, methods and optimized guide compositions with new architectures for sequence manipulation
DK3604544T3 (en) 2014-01-31 2022-02-14 Swift Biosciences Inc Improved methods for processing dna substrates
EP3919621A1 (en) 2014-06-23 2021-12-08 The General Hospital Corporation Genomewide unbiased identification of dsbs evaluated by sequencing (guide-seq)
AU2016206870B2 (en) 2015-01-12 2022-02-17 Massachusetts Institute Of Technology Gene editing through microfluidic delivery
MA41349A (fr) 2015-01-14 2017-11-21 Univ Temple Éradication de l'herpès simplex de type i et d'autres virus de l'herpès associés guidée par arn
IL293323B2 (en) 2015-06-18 2024-01-01 Massachusetts Inst Technology Crispr enzyme mutations reducing off-target effects
US9790490B2 (en) 2015-06-18 2017-10-17 The Broad Institute Inc. CRISPR enzymes and systems
WO2017015015A1 (en) 2015-07-17 2017-01-26 Emory University Crispr-associated protein from francisella and uses related thereto
US9926546B2 (en) 2015-08-28 2018-03-27 The General Hospital Corporation Engineered CRISPR-Cas9 nucleases
US9512446B1 (en) 2015-08-28 2016-12-06 The General Hospital Corporation Engineered CRISPR-Cas9 nucleases
AU2016316845B2 (en) 2015-08-28 2022-03-10 The General Hospital Corporation Engineered CRISPR-Cas9 nucleases
CN108350453A (zh) 2015-09-11 2018-07-31 通用医疗公司 核酸酶dsb的完全查询和测序(find-seq)
JP2018529353A (ja) 2015-09-30 2018-10-11 ザ ジェネラル ホスピタル コーポレイション 配列決定法による切断事象の包括的生体外報告（ＣＩＲＣＬＥ−ｓｅｑ）
EP3845668A1 (en) 2015-10-30 2021-07-07 New England Biolabs, Inc. Compositions and methods for determining modified cytosines by sequencing
AU2016348439B2 (en) 2015-11-04 2023-03-09 Atreca, Inc. Combinatorial sets of nucleic acid barcodes for analysis of nucleic acids associated with single cells
EP3374501B1 (en) 2015-11-11 2023-07-12 Lonza Ltd Crispr-associated (cas) proteins with reduced immunogenicity
US9896696B2 (en) 2016-02-15 2018-02-20 Benson Hill Biosystems, Inc. Compositions and methods for modifying genomes
CN105543195A (zh) * 2016-03-01 2016-05-04 中国水稻研究所 植物Cas9变体蛋白VQR及其编码基因与应用
SG11201810179RA (en) 2016-04-19 2018-12-28 Broad Inst Inc Novel crispr enzymes and systems
CN109312386B (zh) * 2016-06-15 2022-10-25 株式会社图尔金 使用中靶靶标和脱靶靶标的多重靶标系统筛选靶特异性核酸酶的方法及其用途
KR20250103795A (ko) 2016-08-03 2025-07-07 프레지던트 앤드 펠로우즈 오브 하바드 칼리지 아데노신 핵염기 편집제 및 그의 용도
WO2018052247A1 (ko) 2016-09-13 2018-03-22 주식회사 툴젠 시토신 디아미나제에 의한 dna에서의 염기 교정 확인 방법
CA3039409A1 (en) 2016-10-07 2018-04-12 Integrated Dna Technologies, Inc. S. pyogenes cas9 mutant genes and polypeptides encoded by same
US11242542B2 (en) 2016-10-07 2022-02-08 Integrated Dna Technologies, Inc. S. pyogenes Cas9 mutant genes and polypeptides encoded by same
SG10201913505WA (en) 2016-10-17 2020-02-27 Univ Nanyang Tech Truncated crispr-cas proteins for dna targeting
WO2018119359A1 (en) 2016-12-23 2018-06-28 President And Fellows Of Harvard College Editing of ccr5 receptor gene to protect against hiv infection
AU2018273986A1 (en) 2017-05-25 2019-12-12 The General Hospital Corporation Bipartite base editor (BBE) architectures and type-II-C-Cas9 zinc finger editing
CN110914423B (zh) * 2017-05-31 2024-02-06 国立大学法人东京大学 经修饰的Cas9蛋白及其用途
US20200140835A1 (en) 2017-06-06 2020-05-07 The General Hospital Corporation Engineered CRISPR-Cas9 Nucleases
BR112020000310A2 (pt) 2017-07-07 2020-07-14 Toolgen Incorporated variantes de crispr alvo específicas
CA3073448A1 (en) 2017-08-23 2019-02-28 The General Hospital Corporation Engineered crispr-cas9 nucleases with altered pam specificity
WO2019075197A1 (en) 2017-10-11 2019-04-18 The General Hospital Corporation METHODS OF DETECTION OF INDIVIDUAL SITE-SPECIFIC PARASITE GENOMIC DEAMINATION BY BASE EDITING TECHNOLOGIES
EP3707252A1 (en) 2017-11-10 2020-09-16 Novozymes A/S Temperature-sensitive cas9 protein
EP3781585A4 (en) 2018-04-17 2022-01-26 The General Hospital Corporation SENSITIVE IN VITRO TESTS FOR SUBSTRATE PREFERENCES AND SITE FOR NUCLEIC ACID BINDERS, MODIFIERS AND CLEAVAGE AGENTS
KR20250133482A (ko) 2018-05-11 2025-09-05 빔 테라퓨틱스, 인크. 프로그래밍가능한 염기 편집기 시스템을 이용하여 단일 뉴클레오티드 다형성을 편집하는 방법
US20230021641A1 (en) 2018-08-23 2023-01-26 The Broad Institute, Inc. Cas9 variants having non-canonical pam specificities and uses thereof
US20220411777A1 (en) 2019-08-30 2022-12-29 The General Hospital Corporation C-to-G Transversion DNA Base Editors
WO2021151073A2 (en) 2020-01-24 2021-07-29 The General Hospital Corporation Unconstrained genome targeting with near-pamless engineered crispr-cas9 variants
WO2021151085A2 (en) 2020-01-24 2021-07-29 The General Hospital Corporation Crispr-cas enzymes with enhanced on-target activity

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016141224A1 (en) * 2015-03-03 2016-09-09 The General Hospital Corporation Engineered crispr-cas9 nucleases with altered pam specificity
WO2017070633A2 (en) * 2015-10-23 2017-04-27 President And Fellows Of Harvard College Evolved cas9 proteins for gene editing

Also Published As

Publication number Publication date
US12241096B2 (en) 2025-03-04
US11286468B2 (en) 2022-03-29
US20220169998A1 (en) 2022-06-02
US11624058B2 (en) 2023-04-11
BR112020003596A2 (pt) 2020-09-01
AU2018320865A1 (en) 2020-03-05
CN111278450B (zh) 2024-04-09
AU2025275152A1 (en) 2026-01-08
CA3073448A1 (en) 2019-02-28
AU2023278074A1 (en) 2024-01-04
AU2023278074B2 (en) 2025-09-11
AU2025275152B2 (en) 2026-03-12
EP3672612A4 (en) 2021-09-29
JP2020532967A (ja) 2020-11-19
EP3672612A1 (en) 2020-07-01
US20190106687A1 (en) 2019-04-11
CN111278450A (zh) 2020-06-12
US20230407277A1 (en) 2023-12-21
JP7581046B2 (ja) 2024-11-12
WO2019040650A1 (en) 2019-02-28
JP2024112895A (ja) 2024-08-21
CN118530967A (zh) 2024-08-23

Similar Documents

Publication Publication Date Title
AU2023278074B2 (en) Engineered CRISPR-Cas9 nucleases with altered PAM specificity
US12180520B2 (en) Engineered CRISPR-Cas9 nucleases with altered PAM specificity
US20250382595A1 (en) CRISPR-Cas Enzymes with Enhanced On-Target Activity
US12312613B2 (en) Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants
CN110352241A (zh) 热稳定的Cas9核酸酶
HK1249132B (en) Engineered crispr-cas9 nucleases with altered pam specificity

Legal Events

Date Code Title Description
FGA Letters patent sealed or granted (standard patent)