EP4709853A1 - Procédés de prévention de la génotoxicité sur cible induite par des nucléases - Google Patents

Procédés de prévention de la génotoxicité sur cible induite par des nucléases

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Publication number
EP4709853A1
EP4709853A1 EP24723913.0A EP24723913A EP4709853A1 EP 4709853 A1 EP4709853 A1 EP 4709853A1 EP 24723913 A EP24723913 A EP 24723913A EP 4709853 A1 EP4709853 A1 EP 4709853A1
Authority
EP
European Patent Office
Prior art keywords
cells
cell
loh
nuclease
editing
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
EP24723913.0A
Other languages
German (de)
English (en)
Inventor
François MOREAU-GAUDRY
Aurélie BEDEL
Sabrina FAYET
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.)
Institut National de la Sante et de la Recherche Medicale INSERM
Centre Hospitalier Universitaire de Bordeaux
Universite de Bordeaux
Original Assignee
Institut National de la Sante et de la Recherche Medicale INSERM
Centre Hospitalier Universitaire de Bordeaux
Universite de Bordeaux
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 Institut National de la Sante et de la Recherche Medicale INSERM, Centre Hospitalier Universitaire de Bordeaux, Universite de Bordeaux filed Critical Institut National de la Sante et de la Recherche Medicale INSERM
Publication of EP4709853A1 publication Critical patent/EP4709853A1/fr
Pending legal-status Critical Current

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
    • 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
    • C12N15/87—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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or 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
    • 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

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Immunology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Microbiology (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Food Science & Technology (AREA)
  • Plant Pathology (AREA)
  • Pathology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Mycology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Crystallography & Structural Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Le système CRISPR-Cas9 a révolutionné notre aptitude à modifier avec précision le génome et présente une édition de gènes poussée dans des applications cliniques. Une analyse complète de produits d'édition de gènes au niveau du site de coupe ciblé a révélé un spectre complexe de résultats. Une génotoxicité sur cible est sous-estimée avec des procédés basés sur la PCR standard et nécessite des procédés de détection appropriés et sensibles. Ici, les inventeurs ont développé deux systèmes de détection de réarrangements à l'échelle de la mégabase assistés par fluorescence (FAMReD) complémentaires qui garantissent la détection, la quantification et le tri cellulaire de cellules éditées avec une perte d'hétérozygosité (LOH) à l'échelle de la mégabase. Ils ont révélé des réarrangements chromosomiques complexes rares provoqués par la nucléase Cas9 et ont montré que la fréquence LOH dépend du taux de division cellulaire lors de l'édition et de l'état p53. L'arrêt du cycle cellulaire lors de l'édition a supprimé l'apparition de LOH sans compromettre l'édition. Ces données ont été confirmées dans des cellules souches/progénitrices humaines, suggérant que des essais cliniques doivent tenir compte de l'état p53 et du taux de prolifération cellulaire lors de l'édition pour limiter ce risque et concevoir des protocoles plus sûrs.
EP24723913.0A 2023-05-12 2024-05-10 Procédés de prévention de la génotoxicité sur cible induite par des nucléases Pending EP4709853A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
EP23305760 2023-05-12
PCT/EP2024/062916 WO2024235844A1 (fr) 2023-05-12 2024-05-10 Procédés de prévention de la génotoxicité sur cible induite par des nucléases

Publications (1)

Publication Number Publication Date
EP4709853A1 true EP4709853A1 (fr) 2026-03-18

Family

ID=86760357

Family Applications (1)

Application Number Title Priority Date Filing Date
EP24723913.0A Pending EP4709853A1 (fr) 2023-05-12 2024-05-10 Procédés de prévention de la génotoxicité sur cible induite par des nucléases

Country Status (2)

Country Link
EP (1) EP4709853A1 (fr)
WO (1) WO2024235844A1 (fr)

Family Cites Families (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040006074A1 (en) 1998-04-28 2004-01-08 The Government Of The United States Of America Cyclin dependent kinase (CDK)4 inhibitors and their use for treating cancer
ES2251677T3 (es) 2002-01-22 2006-05-01 Warner-Lambert Company Llc 2-(piridin-2-ilamino)-pirido(2,3-d)pirimidin-7-onas.
ES2522346T3 (es) 2008-08-22 2014-11-14 Novartis Ag Compuestos de pirrolopirimidina como inhibidores de CDK
JO2885B1 (en) 2008-12-22 2015-03-15 ايلي ليلي اند كومباني Protein kinase inhibitors
CA2963820A1 (fr) * 2014-11-07 2016-05-12 Editas Medicine, Inc. Procedes pour ameliorer l'edition genomique mediee par crispr/cas
WO2019241802A2 (fr) * 2018-06-15 2019-12-19 Ideaya Biosciences, Inc. Procédés d'inhibition de cellules prolifératives

Also Published As

Publication number Publication date
WO2024235844A1 (fr) 2024-11-21

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