【摘 要】
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Dear Editor,The clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) system is an adaptive immune system in a variety of bacteria
【机 构】
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State Key Laboratory of Stem Cell and Reproductive Biology,Institute of Zoology, Chinese Academy of
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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 (Tes and Tes,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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