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Tytuł:
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CRISPR-Cas9 globin editing can induce megabase-scale copy-neutral losses of heterozygosity in hematopoietic cells.
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Autorzy:
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Boutin J; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Rosier J; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Cappellen D; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.; University Hospital Bordeaux, Tumor Biology and Tumor Bank Laboratory, Bordeaux, France.
Prat F; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Toutain J; Bordeaux University, MRGM INSERM U1211, CHU de Bordeaux, Service de Génétique Médicale, Bordeaux, France.
Pennamen P; Bordeaux University, MRGM INSERM U1211, CHU de Bordeaux, Service de Génétique Médicale, Bordeaux, France.
Bouron J; Bordeaux University, MRGM INSERM U1211, CHU de Bordeaux, Service de Génétique Médicale, Bordeaux, France.
Rooryck C; Bordeaux University, Bordeaux, France.; Bordeaux University, MRGM INSERM U1211, CHU de Bordeaux, Service de Génétique Médicale, Bordeaux, France.
Merlio JP; Bordeaux University, Bordeaux, France.; University Hospital Bordeaux, Tumor Biology and Tumor Bank Laboratory, Bordeaux, France.; INSERM U1053, Bordeaux Research in Translational Oncology, Bordeaux, France.
Lamrissi-Garcia I; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Cullot G; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Amintas S; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.; University Hospital Bordeaux, Tumor Biology and Tumor Bank Laboratory, Bordeaux, France.
Guyonnet-Duperat V; INSERM US 005-CNRS UMS 342-TBM-Core, Bordeaux University, Bordeaux, France.
Ged C; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Blouin JM; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Richard E; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Dabernat S; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
Moreau-Gaudry F; Bordeaux University, Bordeaux, France. .; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France. .; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France. .; Laboratory of Excellence, Gr-Ex, Bordeaux, France. .; INSERM US 005-CNRS UMS 342-TBM-Core, Bordeaux University, Bordeaux, France. .
Bedel A; Bordeaux University, Bordeaux, France.; INSERM U1035, Biotherapy of Genetic Diseases, Inflammatory disorders and Cancers, Bordeaux, France.; University Hospital Bordeaux, Biochemistry Laboratory, Bordeaux, France.; Laboratory of Excellence, Gr-Ex, Bordeaux, France.
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Źródło:
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Nature communications [Nat Commun] 2021 Aug 13; Vol. 12 (1), pp. 4922. Date of Electronic Publication: 2021 Aug 13.
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Typ publikacji:
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Journal Article; Research Support, Non-U.S. Gov't
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Język:
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English
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Imprint Name(s):
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Original Publication: [London] : Nature Pub. Group
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MeSH Terms:
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CRISPR-Cas Systems*
Sequence Deletion*
Gene Editing/*methods
Globins/*genetics
Hematopoietic Stem Cells/*metabolism
Loss of Heterozygosity/*genetics
Cells, Cultured ; Chromosome Deletion ; Chromosomes, Human, Pair 11/genetics ; DNA Methylation ; Gene Expression ; HEK293 Cells ; Hematopoietic Stem Cells/cytology ; Humans ; Insulin-Like Growth Factor II/genetics ; Polymorphism, Single Nucleotide ; RNA, Long Noncoding/genetics
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Substance Nomenclature:
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0 (H19 long non-coding RNA)
0 (RNA, Long Noncoding)
67763-97-7 (Insulin-Like Growth Factor II)
9004-22-2 (Globins)
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Entry Date(s):
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Date Created: 20210814 Date Completed: 20210830 Latest Revision: 20231107
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Update Code:
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20240105
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PubMed Central ID:
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PMC8363739
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DOI:
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10.1038/s41467-021-25190-6
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PMID:
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34389729
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CRISPR-Cas9 is a promising technology for gene therapy. However, the ON-target genotoxicity of CRISPR-Cas9 nuclease due to DNA double-strand breaks has received little attention and is probably underestimated. Here we report that genome editing targeting globin genes induces megabase-scale losses of heterozygosity (LOH) from the globin CRISPR-Cas9 cut-site to the telomere (5.2 Mb). In established lines, CRISPR-Cas9 nuclease induces frequent terminal chromosome 11p truncations and rare copy-neutral LOH. In primary hematopoietic progenitor/stem cells, we detect 1.1% of clones (7/648) with acquired megabase LOH induced by CRISPR-Cas9. In-depth analysis by SNP-array reveals the presence of copy-neutral LOH. This leads to 11p15.5 partial uniparental disomy, comprising two Chr11p15.5 imprinting centers (H19/IGF2:IG-DMR/IC1 and KCNQ1OT1:TSS-DMR/IC2) and impacting H19 and IGF2 expression. While this genotoxicity is a safety concern for CRISPR clinical trials, it is also an opportunity to model copy-neutral-LOH for genetic diseases and cancers.
(© 2021. The Author(s).)
