학술논문
An efficient KRAB domain for CRISPRi applications in human cells
clustered regularly interspaced short palindromic repeat interference
clustered regularly interspaced short palindromic repeat interference
Document Type
Report
Source
Nature Methods. November 2020, Vol. 17 Issue 11, p1093, 4 p.
Subject
Language
English
ISSN
1548-7091
Abstract
Author(s): Nader Alerasool [sup.1] [sup.2] , Dmitri Segal [sup.1] [sup.2] , Hunsang Lee [sup.1] , Mikko Taipale [sup.1] [sup.2] [sup.3] Author Affiliations: (1) Donnelly Centre for Cellular and Biomolecular Research, [...]
Clustered regularly interspaced short palindromic repeat interference (CRISPRi), based on the fusion of inactive Cas9 (dCas9) to the Krüppel-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. 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 Krüppel-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. 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.