Retained chromosomal integrity following CRISPR-Cas9-based mutational correction in human embryos.
CRISPR-Cas9
chromosomal integrity
gene conversion
germline genome editing
human embryo
male infertility
mosaicism
Journal
Molecular therapy : the journal of the American Society of Gene Therapy
ISSN: 1525-0024
Titre abrégé: Mol Ther
Pays: United States
ID NLM: 100890581
Informations de publication
Date de publication:
02 08 2023
02 08 2023
Historique:
received:
09
12
2022
revised:
11
04
2023
accepted:
22
06
2023
pmc-release:
02
08
2024
medline:
7
8
2023
pubmed:
28
6
2023
entrez:
28
6
2023
Statut:
ppublish
Résumé
Human germline gene correction by targeted nucleases holds great promise for reducing mutation transmission. However, recent studies have reported concerning observations in CRISPR-Cas9-targeted human embryos, including mosaicism and loss of heterozygosity (LOH). The latter has been associated with either gene conversion or (partial) chromosome loss events. In this study, we aimed to correct a heterozygous basepair substitution in PLCZ1, related to infertility. In 36% of the targeted embryos that originated from mutant sperm, only wild-type alleles were observed. By performing genome-wide double-digest restriction site-associated DNA sequencing, integrity of the targeted chromosome (i.e., no deletions larger than 3 Mb or chromosome loss) was confirmed in all seven targeted GENType-analyzed embryos (mutant editing and absence of mutation), while short-range LOH events (shorter than 10 Mb) were clearly observed by single-nucleotide polymorphism assessment in two of these embryos. These results fuel the currently ongoing discussion on double-strand break repair in early human embryos, making a case for the occurrence of gene conversion events or partial template-based homology-directed repair.
Identifiants
pubmed: 37376733
pii: S1525-0016(23)00324-6
doi: 10.1016/j.ymthe.2023.06.013
pmc: PMC10422011
pii:
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2326-2341Informations de copyright
Copyright © 2023 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests D.S. is a member of the board of the BSRM (Belgian Society for Reproductive Medicine) and chairman of the Belgian College for reproductive medicine. Additionally, D.S. is a member of the advisory board of Organon Pharmaceuticals and receives honoraria for lectures from Organon, Ferring, and Gedeon Richter. B.H. is the vice president of BSRM.
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