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An enhanced CRISPR repressor for targeted mammalian gene regulation
Nature Methods ( IF 48.0 ) Pub Date : 2018-07-16 , DOI: 10.1038/s41592-018-0048-5
Nan Cher Yeo , Alejandro Chavez , Alissa Lance-Byrne , Yingleong Chan , David Menn , Denitsa Milanova , Chih-Chung Kuo , Xiaoge Guo , Sumana Sharma , Angela Tung , Ryan J. Cecchi , Marcelle Tuttle , Swechchha Pradhan , Elaine T. Lim , Noah Davidsohn , Mo R. Ebrahimkhani , James J. Collins , Nathan E. Lewis , Samira Kiani , George M. Church

The RNA-guided endonuclease Cas9 can be converted into a programmable transcriptional repressor, but inefficiencies in target-gene silencing have limited its utility. Here we describe an improved Cas9 repressor based on the C-terminal fusion of a rationally designed bipartite repressor domain, KRAB–MeCP2, to nuclease-dead Cas9. We demonstrate the system’s superiority in silencing coding and noncoding genes, simultaneously repressing a series of target genes, improving the results of single and dual guide RNA library screens, and enabling new architectures of synthetic genetic circuits.



中文翻译:

用于靶定哺乳动物基因调控的增强型CRISPR阻遏物

RNA引导的核酸内切酶Cas9可以转化为可编程的转录阻遏物,但是靶基因沉默的低效率限制了其实用性。在这里,我们描述了一种经过改进的Cas9阻遏物,其基于合理设计的两部分阻遏物结构域KRAB-MeCP2与核酸酶死亡的Cas9的C端融合。我们展示了该系统在沉默编码和非编码基因,同时抑制一系列靶基因,改善单和双指导RNA库筛选结果以及实现合成遗传电路新架构方面的优越性。

更新日期:2018-07-18
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