Enhancing homology-directed repair efficiency with HDR-boosting modular ssDNA donor.
Nat Commun
; 15(1): 6843, 2024 Aug 10.
Article
em En
| MEDLINE
| ID: mdl-39122671
ABSTRACT
Despite the potential of small molecules and recombinant proteins to enhance the efficiency of homology-directed repair (HDR), single-stranded DNA (ssDNA) donors, as currently designed and chemically modified, remain suboptimal for precise gene editing. Here, we screen the biased ssDNA binding sequences of DNA repair-related proteins and engineer RAD51-preferred sequences into HDR-boosting modules for ssDNA donors. Donors with these modules exhibit an augmented affinity for RAD51, thereby enhancing HDR efficiency across various genomic loci and cell types when cooperated with Cas9, nCas9, and Cas12a. By combining with an inhibitor of non-homologous end joining (NHEJ) or the HDRobust strategy, these modular ssDNA donors achieve up to 90.03% (median 74.81%) HDR efficiency. The HDR-boosting modules targeting an endogenous protein enable a chemical modification-free strategy to improve the efficacy of ssDNA donors for precise gene editing.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
DNA de Cadeia Simples
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Rad51 Recombinase
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Reparo de DNA por Recombinação
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Edição de Genes
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article