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Diverse noncanonical PAMs recognized by SpCas9 in human cells

Hu, Z.; Wang, D.; Zhang, C.; Wang, S.; Gao, S.; Hou, L.; Wang, H.; Wang, Y.

2019-06-14 bioengineering
10.1101/671503 bioRxiv
Show 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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