Journal
NUCLEIC ACIDS RESEARCH
Volume 46, Issue 6, Pages 2945-2955Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gky076
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Funding
- Netherlands Organization for Health Research and Development [ZonMw-TOP 40-00812-98-10033]
- Netherlands Cancer Institute
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Single-stranded oligodeoxyribonucleotide (ssODN)-mediated repair of CRISPR/Cas9-induced DNA double-strand breaks (DSB) can effectively be used to introduce small genomic alterations in a defined locus. Here, we reveal DNA mismatch repair (MMR) activity is crucial for efficient nucleotide substitution distal from the Cas9-induced DNA break when the substitution is instructed by the 3' half of the ssODN. Furthermore, protecting the ssODN 3' end with phosphorothioate linkages enhances MMR-dependent gene editing events. Our findings can be exploited to optimize efficiencies of nucleotide substitutions distal from the DSB and imply that oligonucleotide-mediated gene editing is effectuated by templated break repair.
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