Regulation of transcriptionally active genes via the catalytically inactive Cas9 in C.elegans and D.rerio
Dear Editor,CRISPR interference (CRISPRi) is a recently developed tool used to study single guide RNA (gRNA)-mediated sequence-specific repression of transcription in both prokaryotic and eukaryotic cells ”1-5”.Transcription initiation and elongation of a gene can be interfered by the presence of gRNA:DNA hetero-duplex/dCas9 (a catalytically inactive form of Cas9) complex in its promoter and exons.If Krüppel associated box (KRAB) domain is fused to dCas9,repression of target gene is more efficient.Likewise,fusion of a transcription activator such as VP16 (CRISPR-on) can increase target gene expression through three to four gene-specific gRNAs (up to seven) that recognize the proximal promoter of a target gene in cultured cells and in vivo ”2”.We applied both CRISPRi and CRISPR-on tools in worm and zebrafish,and demonstrated here that our dCas9 fusion systems modify gene expression at or near their endogenous expression location(s) through target-specific gRNAs (ts-gRNAs).
25
This study was supported by the National Natural Science Foundation of China 31371316,the National Basic Research Program of China 973 Program;2012CB944503 and 2011CBA01102,the Ministry of Science and Technology of China and the Peking-Tsinghua Center for Life Sciences to DL.
2015-06-10(万方平台首次上网日期,不代表论文的发表时间)
638-641