期刊
NATURE COMMUNICATIONS
卷 10, 期 -, 页码 -出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/s41467-018-07988-z
关键词
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资金
- National Key R&D Program of China [2017YFC1001901, 2017YFA0102801, 2017YFC1001603]
- National Natural Science Foundation [91640119, 31671540, 81330055, 31601196]
- Natural Science Foundation of Guangdong Province [2016A030310206, 2014A030312011]
- Science and Technology Planning Project of Guangdong Province [2015B020228002]
- Guangzhou Science and Technology Project [201707010085, 201803010020]
The adenine base editor (ABE), capable of catalyzing A.T to G.C conversions, is an important gene editing toolbox. Here, we systematically evaluate genome-wide off-target deamination by ABEs using the EndoV-seq platform we developed. EndoV-seq utilizes Endonuclease V to nick the inosine-containing DNA strand of genomic DNA deaminated by ABE in vitro. The treated DNA is then whole-genome sequenced to identify off-target sites. Of the eight gRNAs we tested with ABE, 2-19 (with an average of 8.0) off-target sites are found, significantly fewer than those found for canonical Cas9 nuclease (7-320, 160.7 on average). In vivo off-target deamination is further validated through target site deep sequencing. Moreover, we demonstrated that six different ABE-gRNA complexes could be examined in a single EndoV-seq assay. Our study presents the first detection method to evaluate genome-wide off-target effects of ABE, and reveals possible similarities and differences between ABE and canonical Cas9 nuclease.
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