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论文题目 Shortening the sgRNA-DNA interface enables SpCas9 and eSpCas9 to nick the target DNA strand
作  者 Fan, Rong (范荣); Chai, Zhuangzhuang; Xing, Sinian Chen, Kunling; Qiu, Fengti; Tuanyao Chai; Jin-Long Qiu; Zhang, Zhengbin (张正斌); Zhang, Huawei Gao, Caixia;
发表年度 2020
刊物名称 SCIENCE CHINA Life Sciences
卷、期、页码 published online; ;
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论文摘要
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.
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