Expanding the scope of CRISPR/Cas9-mediated genome editing in plants using an xCas9 and Cas9-NG hybrid^FA  

作　　者：Qingfeng Niu Siqun Wu Yansha Li Xiaoxuan Yang Ping Liu Yaping Xu Zhaobo Lang 

机构地区：[1]Shanghai Center for Plant Stress Biology,and National Key Laboratory of Plant Molecular Genetics,Center of Excellence in Molecular Plant Sciences,Chinese Academy of Sciences,Shanghai 200032,China [2]Shandong Shunfeng Biotechnology Co.,Ltd,Jinan 250000,China

出　　处：《Journal of Integrative Plant Biology》2020年第4期398-402,共5页植物学报（英文版）

基　　金：supported by the National Key Research and Development Program(2018YFD1000200)(Z.L.);Major Project of Transgenic Crops of China(2019ZX08010003-002-006)(Y.L.);Strategic Priority Research Program of the Chinese Academy of Sciences(XDB27040000)(Z.L.)。

摘　　要：The widely used Streptococcus pyogenes Cas9(SpCas9)requires NGG as a protospacer adjacent motif(PAM)for genome editing.Although Sp Cas9 is a powerful genome-editing tool,its use has been limited on the targetable genomic locus lacking NGG PAM.The Sp Cas9 variants xCas9 and Cas9-NG have been developed to recognize NG,GAA,and GAT PAMs in human cel s.Here,we show that xCas9 cannot recognize NG PAMs in tomato,and Cas9-NG can recognize some of our tested NG PAMs in the tomato and Arabidopsis genomes.In addition,we engineered BrSp Cas9(XNG-Cas9)based on mutations from both xCas9 and Cas9-NG,and found that XNG-Cas9 can efficiently mutagenize endogenous target sites with NG,GAG,GAA,and GAT PAMs in the tomato or Arabidopsis genomes.The PAM compatibility of XNG-Cas9 is the broadest reported to date among Cas9 s(SpCas9 and Cas9-NG)active in plant.

关 键 词：CRISPR/Cas9 EDITING SPACER 

分 类 号：Q943.2[生物学—植物学]