IL274528B2 - Engineered Cas9 systems for eukaryotic genome modification - Google Patents

Engineered Cas9 systems for eukaryotic genome modification

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Publication number
IL274528B2
IL274528B2 IL274528A IL27452820A IL274528B2 IL 274528 B2 IL274528 B2 IL 274528B2 IL 274528 A IL274528 A IL 274528A IL 27452820 A IL27452820 A IL 27452820A IL 274528 B2 IL274528 B2 IL 274528B2
Authority
IL
Israel
Prior art keywords
engineered
sequence
cas9 protein
domain
bacillus smithii
Prior art date
Application number
IL274528A
Other languages
English (en)
Hebrew (he)
Other versions
IL274528A (en
IL274528B1 (en
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Sigma Aldrich Co Llc
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 Sigma Aldrich Co Llc filed Critical Sigma Aldrich Co Llc
Publication of IL274528A publication Critical patent/IL274528A/en
Publication of IL274528B1 publication Critical patent/IL274528B1/en
Publication of IL274528B2 publication Critical patent/IL274528B2/en

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    • 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
    • C12N15/87Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
    • C12N15/90Stable introduction of foreign DNA into chromosome
    • C12N15/902Stable introduction of foreign DNA into chromosome using homologous recombination
    • C12N15/907Stable introduction of foreign DNA into chromosome using homologous recombination in mammalian cells
    • 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
    • C12N15/11DNA 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
    • 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
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • 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
    • C12N15/63Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • 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
    • 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]
    • C12N9/222Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes
    • C12N9/226Class 2 CAS enzyme complex, e.g. single CAS protein
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/01Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/09Fusion polypeptide containing a localisation/targetting motif containing a nuclear localisation signal
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/01Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/10Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K2319/00Fusion polypeptide
    • C07K2319/85Fusion polypeptide containing an RNA binding domain
    • 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
    • C12Y301/00Hydrolases acting on ester bonds (3.1)
    • C12Y301/21Endodeoxyribonucleases producing 5'-phosphomonoesters (3.1.21)
    • C12Y301/21001Deoxyribonuclease I (3.1.21.1)

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (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)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
IL274528A 2018-02-15 2019-02-15 Engineered Cas9 systems for eukaryotic genome modification IL274528B2 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US201862631304P 2018-02-15 2018-02-15
US201862720525P 2018-08-21 2018-08-21
PCT/US2019/018335 WO2019161290A1 (en) 2018-02-15 2019-02-15 Engineered cas9 systems for eukaryotic genome modification

Publications (3)

Publication Number Publication Date
IL274528A IL274528A (en) 2020-06-30
IL274528B1 IL274528B1 (en) 2025-06-01
IL274528B2 true IL274528B2 (en) 2025-10-01

Family

ID=65995829

Family Applications (2)

Application Number Title Priority Date Filing Date
IL274528A IL274528B2 (en) 2018-02-15 2019-02-15 Engineered Cas9 systems for eukaryotic genome modification
IL321024A IL321024A (en) 2018-02-15 2019-02-15 Engineered CAS9 Systems For Eukaryotic Genome Modification

Family Applications After (1)

Application Number Title Priority Date Filing Date
IL321024A IL321024A (en) 2018-02-15 2019-02-15 Engineered CAS9 Systems For Eukaryotic Genome Modification

Country Status (11)

Country Link
US (2) US10767193B2 (https=)
EP (1) EP3752607A1 (https=)
JP (3) JP7109547B2 (https=)
KR (3) KR102465067B1 (https=)
CN (1) CN111902536B (https=)
AU (3) AU2019222568B2 (https=)
BR (1) BR112020010479A2 (https=)
CA (2) CA3281020A1 (https=)
IL (2) IL274528B2 (https=)
SG (1) SG11202007382TA (https=)
WO (1) WO2019161290A1 (https=)

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AU2015330699B2 (en) 2014-10-10 2021-12-02 Editas Medicine, Inc. Compositions and methods for promoting homology directed repair
US11111492B2 (en) * 2017-03-06 2021-09-07 Florida State University Research Foundation, Inc. Genome engineering methods using a cytosine-specific Cas9
KR20210045360A (ko) 2018-05-16 2021-04-26 신테고 코포레이션 가이드 rna 설계 및 사용을 위한 방법 및 시스템
EP3823633A4 (en) 2018-06-29 2023-05-03 Editas Medicine, Inc. SYNTHETIC LEAD MOLECULES, COMPOSITIONS AND METHODS RELATED THERETO
US10913941B2 (en) 2019-02-14 2021-02-09 Metagenomi Ip Technologies, Llc Enzymes with RuvC domains
US10947517B2 (en) 2019-02-15 2021-03-16 Sigma-Aldrich Co. Llc CRISPR/Cas fusion proteins and systems
MX2021010559A (es) 2019-03-07 2021-12-15 Univ California Polipéptidos efectores de crispr-cas y métodos de uso de estos.
IL292517A (en) 2019-12-17 2022-06-01 Sigma Aldrich Co Llc Genome editing in Bacteroidetes
GB2594339B (en) * 2020-02-14 2023-02-08 Metagenomi Inc Enzymes with RUVC domains
US20230058352A1 (en) * 2020-03-11 2023-02-23 Sigma-Aldrich Co. Llc High Fidelity SpCas9 Nucleases for Genome Modification
WO2021202568A1 (en) 2020-03-31 2021-10-07 Metagenomi Ip Technologies, Llc Class ii, type ii crispr systems
CN112159801B (zh) * 2020-07-13 2022-11-15 复旦大学 SlugCas9-HF蛋白、含有SlugCas9-HF蛋白的基因编辑系统及应用
KR102679001B1 (ko) * 2020-09-21 2024-06-28 고려대학교 산학협력단 신규의 개량된 염기 편집 또는 교정용 융합단백질 및 이의 용도
US20220145334A1 (en) * 2020-11-06 2022-05-12 Pairwise Plants Services, Inc. Compositions and methods for rna-encoded dna-replacement of alleles
CN113717960B (zh) * 2021-08-27 2023-07-18 电子科技大学 一种新Cas9蛋白、CRISPR-Cas9基因组定向编辑载体及基因组编辑方法
AU2022335499A1 (en) 2021-08-27 2024-02-22 Metagenomi Therapeutics, Inc. Enzymes with ruvc domains
AU2022396533A1 (en) 2021-11-24 2024-05-02 Metagenomi Therapeutics, Inc. Endonuclease systems
CN114657181B (zh) * 2022-04-01 2023-08-25 安徽大学 一种靶向H1.4的sgRNA以及H1.4基因编辑方法
EP4592322A1 (en) * 2022-09-23 2025-07-30 Chuang, Woei-jer Fusion protein targeting intergrin alpha(iib)beta3 and containing tissue plasminogen activator or variant thereof and use thereof

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US20170314015A1 (en) * 2014-03-26 2017-11-02 Editas Medicine, Inc. Crispr/cas-related methods and compositions for treating sickle cell disease

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AU2013266968B2 (en) 2012-05-25 2017-06-29 Emmanuelle CHARPENTIER Methods and compositions for RNA-directed target DNA modification and for RNA-directed modulation of transcription
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Also Published As

Publication number Publication date
KR102465067B1 (ko) 2022-11-10
WO2019161290A1 (en) 2019-08-22
US20190249200A1 (en) 2019-08-15
AU2022200130A1 (en) 2022-02-10
US12297449B2 (en) 2025-05-13
KR20220015502A (ko) 2022-02-08
JP2024161376A (ja) 2024-11-19
AU2024202158A1 (en) 2024-05-02
AU2019222568B2 (en) 2021-10-14
CA3084020A1 (en) 2019-08-22
KR20230022258A (ko) 2023-02-14
US10767193B2 (en) 2020-09-08
IL274528A (en) 2020-06-30
EP3752607A1 (en) 2020-12-23
JP2021505180A (ja) 2021-02-18
AU2022200130B2 (en) 2024-03-07
CA3281020A1 (en) 2026-03-02
KR102494449B1 (ko) 2023-01-31
KR20200121342A (ko) 2020-10-23
CN111902536B (zh) 2024-07-30
JP7109547B2 (ja) 2022-07-29
US20200354752A1 (en) 2020-11-12
BR112020010479A2 (pt) 2020-11-24
CA3084020C (en) 2025-11-25
JP2022122910A (ja) 2022-08-23
JP7559005B2 (ja) 2024-10-01
SG11202007382TA (en) 2020-08-28
AU2019222568A1 (en) 2020-05-21
KR102684890B1 (ko) 2024-07-12
IL321024A (en) 2025-07-01
IL274528B1 (en) 2025-06-01
CN111902536A (zh) 2020-11-06

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