期刊
NUCLEIC ACIDS RESEARCH
卷 47, 期 14, 页码 7402-7417出版社
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkz459
关键词
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资金
- MRC Molecular Hematology Unit Core award [MC_UU_12009/2]
- MRC Unit studentship as part of the MRC Molecular Hematology Unit Core award [MC_UU_12009/2]
- Medical Research Council IMPC Strategic Award [53658]
- National Institute for Health [U42OD011174]
- MRC MHU [MC_UU_12009]
- NIHR Oxford BRC
- MRC HIU
- John Fell Fund [131/030, 101/517]
- EPA fund [CF182, CF170]
- WIMM Strategic Alliance awards [G0902418, MC_UU_12025]
- MRC [Oxford's RCUK Open Access Block Grant]
- MRC [MC_UU_00016/2, MC_UP_1502/1] Funding Source: UKRI
The CRISPR system is widely used in genome editing for biomedical research. Here, using either dual paired Cas9(D10A) nickases or paired Cas9 nuclease we characterize unintended larger deletions at on-target sites that frequently evade common genotyping practices. We found that unintended larger deletions are prevalent at multiple distinct loci on different chromosomes, in cultured cells and mouse embryos alike. We observed a high frequency of microhomologies at larger deletion breakpoint junctions, suggesting the involvement of microhomology-mediated end joining in their generation. In populations of edited cells, the distribution of larger deletion sizes is dependent on proximity to sgRNAs and cannot be predicted by microhomology sequences alone.
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