Compact SchCas9 Recognizes the Simple NNGR PAM
Shuai Wang, Huilin Mao, Linghui Hou, Ziying Hu, Yao Wang, Yao Wang, Tao Qi, Chen Tao, Yuan Yang, Chengdong Zhang, Miaomiao Li, Huihui Liu, Shijun Hu, Renjie Chai, Yongming Wang, Yongming Wang
Abstract
Clustered regularly interspaced short palindromic repeat (CRISPR)/SaCas9 is the most popular tool for in vivo genome editing due to its high efficiency and small genome. The authors previously developed four SaCas9 orthologs as genome-editing tools. Here, to expand the targeting scope, they investigate the diversity of protospacer adjacent motifs (PAMs) by screening a list of 16 SaCas9 orthologs, twelve of which display editing activity in mammalian cells. They recognize five types of PAMs: NNGRRT, NNGRRR, NNGRC, NNGA, and NNGR. Importantly, SchCas9 recognizes the simple NNGR PAM, representing the most relaxed PAM preference of compact Cas9s to date. It is further demonstrated that SchCas9 enables efficient genome editing in multiple human cell lines. Altogether, these compact Cas9 tools offer a new option for both basic research and clinical applications.