文献类型：期刊文献

英文题名：Compact SchCas9 Recognizes the Simple NNGR PAM

作者：Wang, Shuai[1] Mao, Huilin[1] Hou, Linghui[1] Hu, Ziying[1] Wang, Yao[1] Qi, Tao[1] Tao, Chen[1] Yang, Yuan[1] Zhang, Chengdong[1] Li, Miaomiao[1] Liu, Huihui[2] Hu, Shijun[3,4] Chai, Renjie[5,6] Wang, Yongming[1,7]

第一作者：Wang, Shuai

通信作者：Wang, YM[1];Chai, RJ[2];Chai, RJ[3];Wang, YM[4]

机构：[1]Fudan Univ, Zhongshan Hosp, Sch Life Sci, State Key Lab Genet Engn, Shanghai 200438, Peoples R China;[2]Chinese Acad Forestry, Expt Ctr Forestry North China, Beijing 102300, Peoples R China;[3]Soochow Univ, Affiliated Hosp 1, Dept Cardiovasc Surg, Suzhou 215000, Peoples R China;[4]Soochow Univ, Coll Med, State Key Lab Radiat Med & Protect, Inst Cardiovasc Sci,Collaborat Innovat Ctr Hemato, Suzhou 215000, Peoples R China;[5]Southeast Univ, Sch Life Sci & Technol, State Key Lab Bioelect, Jiangsu Prov High Tech Key Lab Biomed Res, Nanjing 210096, Peoples R China;[6]Nantong Univ, CoInnovat Ctr Neuroregenerat, Nantong 226001, Peoples R China;[7]Shanghai Engn Res Ctr Ind Microorganisms, Shanghai 200438, Peoples R China

年份：2022

卷号：9

期号：4

外文期刊名：ADVANCED SCIENCE

收录：;EI(收录号：20214911289648);Scopus(收录号：2-s2.0-85120542811);WOS:【SCI-EXPANDED(收录号:WOS:000727233600001)】；

基金：S.W., H.M., L.H., Z.H., Y.W., C.T., and M.L. performed experiments; C.Z., Y.Y, and T.Q. analyzed the data; S.H. revised the manuscript; H.L., R.C., and Y.W. provide experimental guidance; Y.W. wrote the manuscript. This work was supported by grants from the National Natural Science Foundation of China (82070258, 81870199, 31700571, 82030029, 81970882); the Foundation for Innovative Research Group of the National Natural Science Foundation of China (31521003), Open Research Fund of State Key Laboratory of Genetic Engineering, Fudan University (No. SKLGE-2104), Science and Technology Research Program of Shanghai [19DZ2282100], Natural Science Foundation from Jiangsu Province (BE2019711).

语种：英文

外文关键词：CRISPR; Cas9; genome editing; PAM; SaCas9; SchCas9

摘要：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.