5”capped and 3”polyA-tailed sgRNAs enhance the efficiency of CRISPR-Cas9 system
Dear Editor,The clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) system is an adaptive immune system in a variety of bacteria and archaea (Terns and Terns,2011).The most commonly used Streptococcus pyogenes type Ⅱ CRISPR-Cas9 system consists of Cas9 nuclease and two short RNAs,crRNA and tracrRNA,which can be linked together forming one chimeric single guide RNA (sgRNA) (Jinek et al.,2012).Guided by sgRNA,Cas9-sgRNA complex can generate DNA double strand breaks (DSB) at specific genomic loci (Jinek et al.,2012;Cong et al.,2013;Mali et al.,2013).Cas9 with mutations at the catalytic RuvC and HNH domains loses the endonuclease activity (indicated ”dCas9”),while maintains its DNA binding capability (Gilbert et al.,2013).DCas9 fused with transcription effectors,such as VP64,P65-HSF1 and KRAB can be guided to specific promoters by sgRNA,enabling regulation of gene transcription (Gilbert et al.,2013,2014).
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We would like to thank Junning Wei and Yi Yang Beijing Cord Blood Bank for their help in preparing the cord blood samples.This work was supported by the National Natural Science Foundation of ChinaGrant 31471215;Strategic Priority Research Program of the Chinese Academy of SciencesXDA16010205;National Key Research and Development Program of China2016YFA0101402;National High-tech R&D Program 863 Program No.2015AA020307.H.Wang is supported by the ”Young Thousand Talents Plan”
2019-05-09（万方平台首次上网日期，不代表论文的发表时间）
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