The PROTECTOR strategy employs dCas orthologs to sterically shield off-target sites from CRISPR/Cas activity.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
09 02 2023
09 02 2023
Historique:
received:
22
10
2022
accepted:
02
02
2023
entrez:
9
2
2023
pubmed:
10
2
2023
medline:
14
2
2023
Statut:
epublish
Résumé
Off-target mutagenesis of CRISPR/Cas systems must be solved to facilitate safe gene therapy. Here, we report a novel approach, termed "PROTECTOR", to shield known off-target sites by directing the binding of an orthologous nuclease-dead Cas protein to the off-target site to sterically interfere with Cas activity. We show that this method reduces off-target mutation rates of two well-studied guide RNAs without compromising on-target activity and that it can be used in combination with high-fidelity Cas enzymes to further reduce off-target editing. This expands the suite of off-target mitigation strategies and offers an ability to protect off-target sites even when their sequences are fully identical to target sites.
Identifiants
pubmed: 36759683
doi: 10.1038/s41598-023-29332-2
pii: 10.1038/s41598-023-29332-2
pmc: PMC9911626
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
2280Informations de copyright
© 2023. The Author(s).
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