Inducing multiple nicks promotes interhomolog homologous recombination to correct heterozygous mutations in somatic cells.
Nat Commun
; 14(1): 5607, 2023 09 15.
Article
em En
| MEDLINE
| ID: mdl-37714828
ABSTRACT
CRISPR/Cas9-mediated gene editing has great potential utility for treating genetic diseases. However, its therapeutic applications are limited by unintended genomic alterations arising from DNA double-strand breaks and random integration of exogenous DNA. In this study, we propose NICER, a method for correcting heterozygous mutations that employs multiple nicks (MNs) induced by Cas9 nickase and a homologous chromosome as an endogenous repair template. Although a single nick near the mutation site rarely leads to successful gene correction, additional nicks on homologous chromosomes strongly enhance gene correction efficiency via interhomolog homologous recombination (IH-HR). This process partially depends on BRCA1 and BRCA2, suggesting the existence of several distinct pathways for MN-induced IH-HR. According to a genomic analysis, NICER rarely induces unintended genomic alterations. Furthermore, NICER restores the expression of disease-causing genes in cells derived from genetic diseases with compound heterozygous mutations. Overall, NICER provides a precise strategy for gene correction.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Recombinação Homóloga
/
Antibacterianos
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article