Shortening the sgRNA-DNA interface enables SpCas9 and eSpCas9(1.1)to nick the target DNA strand  

作　　者：Rong Fan Zhuangzhuang Chai Sinian Xing Kunling Chen Fengti Qiu Tuanyao Chai Jin-Long Qiu Zhengbin Zhang Huawei Zhang Caixia Gao 

机构地区：[1]Key Laboratory of Agricultural Water Resources,Hebei Laboratory of Agricultural Water-Saving,Center for Agricultural Resources Research,Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Shijiazhuang 050022,China [2]State Key Laboratory of Plant Cell and Chromosome Engineering,Center for Genome Editing,Institute of Genetics and Developmental Biology,Innovation Academy for Seed Design,Chinese Academy of Sciences,Beijing 100101,China [3]College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing 100049,China [4]College of Life Sciences,University of Chinese Academy of Sciences,Beijing 100049,China [5]State Key Laboratory of Plant Genomics,Institute of Microbiology,Innovation Academy for Seed Design,Chinese Academy of Sciences,Beijing 100101,China

出　　处：《Science China(Life Sciences)》2020年第11期1619-1630,共12页中国科学（生命科学英文版）

基　　金：supported by grants from the Strategic Priority Research Program of the Chinese Academy of Sciences(Precision Seed Design and Breeding,XDA24020102);the National Transgenic Science and Technology Program(2018ZX0801002B);the National Natural Science Foundation of China(31788103 and 31971370);the Chinese Academy of Sciences(QYZDY-SSW-SMC030);the National Key R&D Program of China(2018YFA0900600,2016YFD0100102-11,and 2016YFD0100605)。

摘　　要：The length of the sgRNA-DNA complementary sequence is a key factor influencing the cleavage activity of Streptococcus pyogenes Cas9(SpCas9)and its variants.The detailed mechanism remains unknown.Here,based on in vitro cleavage assays and base editing analysis,we demonstrate that reducing the length of this complementary region can confer nickase activity on SpCas9 and eSpCas9(1.1).We also show that these nicks are made on the target DNA strand.These properties encouraged us to develop a dual-functional system that simultaneously carries out double-strand DNA cleavage and C-to-T base conversions at separate targets.This system provides a novel tool for achieving trait stacking in plants.

关 键 词：SpCas9 eSpCas9(1.1) truncated spacer DSB nickase co-editing 

分 类 号：Q93[生物学—微生物学]