通过CRISPR/CRISPRi系统对微小RNA进行序列特异性抑制。
Sequence-specific inhibition of microRNA via CRISPR/CRISPRi system.
作者信息
Zhao Yicheng, Dai Zhen, Liang Yang, Yin Ming, Ma Kuiying, He Mei, Ouyang Hongsheng, Teng Chun-Bo
机构信息
1] College of life science, Northeast Forestry University, Harbin, China [2] Department of Biochemistry, College of Animal Sciences, Jilin University, China [3].
1] Department of Biochemistry, College of Animal Sciences, Jilin University, China [2].
出版信息
Sci Rep. 2014 Feb 3;4:3943. doi: 10.1038/srep03943.
Here, we report a convenient and efficient miRNA inhibition strategy employing the CRISPR system. Using specifically designed gRNAs, miRNA gene has been cut at a single site by Cas9, resulting in knockdown of the miRNA in murine cells. Using a modified CRISPR interference system (CRISPRi), inactive Cas9 can reversibly prevent the expression of both monocistronic miRNAs and polycistronic miRNA clusters. Furthermore, CRISPR/CRISPRi is also capable of suppressing genes in porcine cells.
在此，我们报告了一种利用CRISPR系统的便捷高效的miRNA抑制策略。通过使用专门设计的引导RNA（gRNA），Cas9已在单个位点切割miRNA基因，导致小鼠细胞中miRNA的表达下调。使用改良的CRISPR干扰系统（CRISPRi），无活性的Cas9可以可逆地阻止单顺反子miRNA和多顺反子miRNA簇的表达。此外，CRISPR/CRISPRi也能够在猪细胞中抑制基因。
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