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Diverse noncanonical PAMs recognized by SpCas9 in human cells

Ziying Hu, Daqi Wang, Chengdong Zhang, Shuai Wang, Siqi Gao, Linghui Hou, Hongyan Wang, Yongming Wang
doi: https://doi.org/10.1101/671503
Ziying Hu
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Daqi Wang
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Chengdong Zhang
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Shuai Wang
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Siqi Gao
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Linghui Hou
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Hongyan Wang
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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Yongming Wang
1Obstetrics and Gynecology Hospital, State Key Laboratory of Genetic Engineering at School of Life Sciences, Zhongshan Hospital, Fudan University, Shanghai 200011, China
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  • For correspondence: ymw@fudan.edu.cn
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Abstract

The CRISPR/Cas9 system derived from Streptococcus pyogenes (SpCas9) provides unprecedented genome editing capabilities, but the potential for off-target mutations limits its application. In addition to NGG protospacer adjacent motif (PAM), off-target mutations are also associated with noncanonical PAMs, which have not yet been systematically evaluated. Here, we developed a highly sensitive approach that allows systematically analyzing PAM sequences in human cells, and identified multiple alternative PAMs recognized by SpCas9.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted June 14, 2019.
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Diverse noncanonical PAMs recognized by SpCas9 in human cells
Ziying Hu, Daqi Wang, Chengdong Zhang, Shuai Wang, Siqi Gao, Linghui Hou, Hongyan Wang, Yongming Wang
bioRxiv 671503; doi: https://doi.org/10.1101/671503
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Diverse noncanonical PAMs recognized by SpCas9 in human cells
Ziying Hu, Daqi Wang, Chengdong Zhang, Shuai Wang, Siqi Gao, Linghui Hou, Hongyan Wang, Yongming Wang
bioRxiv 671503; doi: https://doi.org/10.1101/671503

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