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WO2024227911A3 - Éditeurs de base crispr hautement actifs obtenus par évolution dirigée liée à un substrat assistée par cas (caslide) - Google Patents

Éditeurs de base crispr hautement actifs obtenus par évolution dirigée liée à un substrat assistée par cas (caslide) Download PDF

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Publication number
WO2024227911A3
WO2024227911A3 PCT/EP2024/062227 EP2024062227W WO2024227911A3 WO 2024227911 A3 WO2024227911 A3 WO 2024227911A3 EP 2024062227 W EP2024062227 W EP 2024062227W WO 2024227911 A3 WO2024227911 A3 WO 2024227911A3
Authority
WO
WIPO (PCT)
Prior art keywords
cas
caslide
highly active
directed evolution
dna
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
PCT/EP2024/062227
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English (en)
Other versions
WO2024227911A2 (fr
Inventor
Duran SÜRÜN
Frank Buchholz
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.)
Technische Universitaet Dresden
Original Assignee
Technische Universitaet Dresden
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
Priority claimed from EP23171650.7A external-priority patent/EP4458963A1/fr
Application filed by Technische Universitaet Dresden filed Critical Technische Universitaet Dresden
Publication of WO2024227911A2 publication Critical patent/WO2024227911A2/fr
Publication of WO2024227911A3 publication Critical patent/WO2024227911A3/fr
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

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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
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1058—Directional evolution of libraries, e.g. evolution of libraries is achieved by mutagenesis and screening or selection of mixed population of organisms
    • 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
    • C12N9/14—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
    • 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
    • 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
    • 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]

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical 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)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Mycology (AREA)
  • Cell Biology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • Ecology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

L'invention concerne une enzyme de modification d'ADN comprenant une protéine d'édition d'ADN et une protéine Cas et un lieur peptidique, ou en étant constitués, ladite protéine d'édition d'ADN et ladite protéine Cas étant reliées par ledit lieur peptidique ; et ladite enzyme de modification d'ADN ayant été obtenue par évolution moléculaire dirigée d'une enzyme d'origine ; et ladite enzyme de modification d'ADN présentant un taux d'édition de base qui est au moins 4 fois supérieur au taux d'édition de base de ladite enzyme de type sauvage. L'invention concerne en outre un procédé d'évolution moléculaire dirigée d'une enzyme de modification d'ADN et l'utilisation d'enzymes de modification d'ADN évoluées dans la recherche, la médecine et l'agriculture.
PCT/EP2024/062227 2023-05-04 2024-05-03 Éditeurs de base crispr hautement actifs obtenus par évolution dirigée liée à un substrat assistée par cas (caslide) Pending WO2024227911A2 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP23171650.7 2023-05-04
EP23171650.7A EP4458963A1 (fr) 2023-05-04 2023-05-04 Éditeurs de base crispr hautement actifs obtenus par évolution dirigée liée à un substrat assistée par cas (caglousier)
EP23194637 2023-08-31
EP23194637.7 2023-08-31

Publications (2)

Publication Number Publication Date
WO2024227911A2 WO2024227911A2 (fr) 2024-11-07
WO2024227911A3 true WO2024227911A3 (fr) 2024-12-12

Family

ID=91027427

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2024/062227 Pending WO2024227911A2 (fr) 2023-05-04 2024-05-03 Éditeurs de base crispr hautement actifs obtenus par évolution dirigée liée à un substrat assistée par cas (caslide)

Country Status (1)

Country Link
WO (1) WO2024227911A2 (fr)

Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20200172931A1 (en) * 2017-07-28 2020-06-04 President And Fellows Of Harvard College Methods and compositions for evolving base editors using phage-assisted continuous evolution (pace)
WO2020181195A1 (fr) * 2019-03-06 2020-09-10 The Broad Institute, Inc. Édition de base t : a à a : t par excision d'adénine
WO2021042062A2 (fr) * 2019-08-30 2021-03-04 Joung J Keith Éditeurs combinatoires d'adénine et de cytosine à base d'adn
CA3198422A1 (fr) * 2020-10-08 2022-04-14 Genkore Inc. Arn guide modifie comprenant une queue riche en u pour augmenter l'efficacite d'un systeme crispr/cas12f1 et utilisation correspondante
WO2022204574A1 (fr) * 2021-03-26 2022-09-29 Beam Therapeutics Inc. Variants de l'adénosine désaminase et leurs utilisations
WO2023282597A1 (fr) * 2021-07-05 2023-01-12 주식회사 진코어 Cas12f1 à clivage inactif, protéine de fusion à base de cas12f1 à clivage inactif, système d'édition génique crispr les comprenant, procédé de préparation et utilisation de ceux-ci
WO2023034959A2 (fr) * 2021-09-03 2023-03-09 The University Of Chicago Polypeptides et procédés de modification d'acides nucléiques

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20200172931A1 (en) * 2017-07-28 2020-06-04 President And Fellows Of Harvard College Methods and compositions for evolving base editors using phage-assisted continuous evolution (pace)
WO2020181195A1 (fr) * 2019-03-06 2020-09-10 The Broad Institute, Inc. Édition de base t : a à a : t par excision d'adénine
WO2021042062A2 (fr) * 2019-08-30 2021-03-04 Joung J Keith Éditeurs combinatoires d'adénine et de cytosine à base d'adn
CA3198422A1 (fr) * 2020-10-08 2022-04-14 Genkore Inc. Arn guide modifie comprenant une queue riche en u pour augmenter l'efficacite d'un systeme crispr/cas12f1 et utilisation correspondante
WO2022204574A1 (fr) * 2021-03-26 2022-09-29 Beam Therapeutics Inc. Variants de l'adénosine désaminase et leurs utilisations
WO2023282597A1 (fr) * 2021-07-05 2023-01-12 주식회사 진코어 Cas12f1 à clivage inactif, protéine de fusion à base de cas12f1 à clivage inactif, système d'édition génique crispr les comprenant, procédé de préparation et utilisation de ceux-ci
WO2023034959A2 (fr) * 2021-09-03 2023-03-09 The University Of Chicago Polypeptides et procédés de modification d'acides nucléiques

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
POPA SERBAN C. ET AL: "Phage-Assisted Continuous Evolution (PACE): A Guide Focused on Evolving Protein-DNA Interactions", ACS OMEGA, vol. 5, no. 42, 16 October 2020 (2020-10-16), US, pages 26957 - 26966, XP055975664, ISSN: 2470-1343, Retrieved from the Internet <URL:http://pubs.acs.org/doi/pdf/10.1021/acsomega.0c03508> DOI: 10.1021/acsomega.0c03508 *
RICHTER MICHELLE F ET AL: "Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity", NATURE BIOTECHNOLOGY, NATURE PUBLISHING GROUP US, NEW YORK, vol. 38, no. 7, 16 March 2020 (2020-03-16), pages 883 - 891, XP037523981, ISSN: 1087-0156, [retrieved on 20200316], DOI: 10.1038/S41587-020-0453-Z *

Also Published As

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