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• W4323348543 abstract "CRISPR-Cas9 introduces targeted DNA breaks that engage competing DNA repair pathways, producing a spectrum of imprecise insertion/deletion mutations (indels) and precise templated mutations (precise edits). The relative frequencies of these pathways are thought to primarily depend on genomic sequence and cell state contexts, limiting control over mutational outcomes. Here, we report that engineered Cas9 nucleases that create different DNA break structures engage competing repair pathways at dramatically altered frequencies. We accordingly designed a Cas9 variant (vCas9) that produces breaks which suppress otherwise dominant nonhomologous end-joining (NHEJ) repair. Instead, breaks created by vCas9 are predominantly repaired by pathways utilizing homologous sequences, specifically microhomology-mediated end-joining (MMEJ) and homology-directed repair (HDR). Consequently, vCas9 enables efficient precise editing through HDR or MMEJ while suppressing indels caused by NHEJ in dividing and nondividing cells. These findings establish a paradigm of targeted nucleases custom-designed for specific mutational applications." @default.
• W4323348543 created "2023-03-08" @default.
• W4323348543 creator A5022945869 @default.
• W4323348543 creator A5042597057 @default.
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• W4323348543 date "2023-03-07" @default.
• W4323348543 modified "2023-09-26" @default.
• W4323348543 title "Altered DNA repair pathway engagement by engineered CRISPR-Cas9 nucleases" @default.
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• W4323348543 doi "https://doi.org/10.1073/pnas.2300605120" @default.
• W4323348543 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/36881621" @default.
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