Journal
NATURE STRUCTURAL & MOLECULAR BIOLOGY
Volume 25, Issue 1, Pages 45-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/s41594-017-0004-6
Keywords
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Funding
- MOST [2014CB910600]
- NSFC [91540115, 31571372, 31471241, 31600619, 31600654]
- Shanghai Pujiang program [16PJ1407000 ', 16PJ1407500]
- CAS Key Laboratory of Computational Biology grants [2015KLCB01, 2016KLCB01]
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The APOBEC-AID family of cytidine deaminase prefers single-stranded nucleic acids for cytidine-to-uridine deamination. Single-stranded nucleic acids are commonly involved in the DNA repair system for breaks generated by CRISPR-Cas9. Here, we show in human cells that APOBEC3 can trigger cytidine deamination of single-stranded oligodeoxynucleotides, which ultimately results in base substitution mutations in genomic DNA through homology-directed repair (HDR) of Cas9-generated double-strand breaks. In addition, the APOBEC3-catalyzed deamination in genomic single-stranded DNA formed during the repair of Cas9 nickase-generated single-strand breaks in human cells can be further processed to yield mutations mainly involving insertions or deletions (indels). Both APOBEC3-mediated deamination and DNA-repair proteins play important roles in the generation of these indels. Therefore, optimizing conditions for the repair of CRISPR-Cas9-generated DNA breaks, such as using double-stranded donors in HDR or temporarily suppressing endogenous APOBEC3s, can repress these unwanted mutations in genomic DNA.
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