Compact SchCas9 Recognizes the Simple NNGR PAM. Issue 4 (6th December 2021)

• Record Type: Journal Article
• Authors: Wang, Shuai; Mao, Huilin; Hou, Linghui; Hu, Ziying; Wang, Yao; Qi, Tao; Tao, Chen; Yang, Yuan; Zhang, Chengdong; Li, Miaomiao; Liu, Huihui; Hu, Shijun; Chai, Renjie; Wang, Yongming
• Is Part Of: Advanced science. Volume 9:Issue 4(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-12-06
• Subjects: CRISPR/Cas9 -- genome editing -- PAM -- SaCas9 -- SchCas9; Science -- Periodicals; 505
• DOI: 10.1002/advs.202104789
• Languages: English
• ISSNs: 2198-3844

Abstract

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. Abstract : The compact clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR‐associated protein‐9 (Cas9) system is a promising platform for therapeutic applications, but its targeting scope is largely restricted to the protospacer‐adjacent motif (PAM). To expand the targeting scope, a Cas9 (SchCas9) recognizing a simple NNGR (R = G or A) PAM is identified. SchCas9 enables efficient genome editing in multiple human cell lines.