CRISPR-Cas9 induces large structural variants at on-target and off-target sites in vivo that segregate across generations.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
02 02 2022
02 02 2022
Historique:
received:
08
10
2021
accepted:
04
01
2022
entrez:
3
2
2022
pubmed:
4
2
2022
medline:
16
2
2022
Statut:
epublish
Résumé
CRISPR-Cas9 genome editing has potential to cure diseases without current treatments, but therapies must be safe. Here we show that CRISPR-Cas9 editing can introduce unintended mutations in vivo, which are passed on to the next generation. By editing fertilized zebrafish eggs using four guide RNAs selected for off-target activity in vitro, followed by long-read sequencing of DNA from >1100 larvae, juvenile and adult fish across two generations, we find that structural variants (SVs), i.e., insertions and deletions ≥50 bp, represent 6% of editing outcomes in founder larvae. These SVs occur both at on-target and off-target sites. Our results also illustrate that adult founder zebrafish are mosaic in their germ cells, and that 26% of their offspring carries an off-target mutation and 9% an SV. Hence, pre-testing for off-target activity and SVs using patient material is advisable in clinical applications, to reduce the risk of unanticipated effects with potentially large implications.
Identifiants
pubmed: 35110541
doi: 10.1038/s41467-022-28244-5
pii: 10.1038/s41467-022-28244-5
pmc: PMC8810904
doi:
Substances chimiques
RNA, Guide
0
DNA
9007-49-2
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
627Informations de copyright
© 2022. The Author(s).
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