An efficient KRAB domain for CRISPRi applications in human cells
An efficient KRAB domain for CRISPRi applications in human cells
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DOI：
10.1038/s41592-020-0966-x
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发表时间：
2020-10-05
期刊：
影响因子：
48
通讯作者：
Taipale, Mikko
中科院分区：
文献类型：
--
作者：
Alerasool, Nader;Segal, Dmitri;Taipale, Mikko
从含有57个KRAB结构域的dCas9融合中测量CRISPRi活性，从而确定ZIM3KRAB-dCas9融合是一个优越的阻遏因子。基于非活性Cas9(DCas9)和Kruppel相关盒(KRAB)阻遏因子的聚集性规则间隔短回文重复干扰(CRISPRi)是一个强大的沉默基因表达的平台。然而，它受到靶基因不完全沉默的影响。我们分析了57个KRAB结构域的抑制活性，并确定ZIM3KRAB结构域是一个特别有效的抑制因子。我们发现，ZIM3KRAB-dCas9融合比现有平台更有效地沉默基因表达。
CRISPRi activities are measured from dCas9 fusions with 57 KRAB domains, which lead to the identification of ZIM3 KRAB-dCas9 fusion as a superior repressor.Clustered regularly interspaced short palindromic repeat interference (CRISPRi), based on the fusion of inactive Cas9 (dCas9) to the Kruppel-associated box (KRAB) repressor, is a powerful platform for silencing gene expression. However, it suffers from incomplete silencing of target genes. We assayed 57 KRAB domains for their repressive potency and identified the ZIM3 KRAB domain as an exceptionally potent repressor. We establish that ZIM3 KRAB-dCas9 fusion silences gene expression more efficiently than existing platforms.