An efficient KRAB domain for CRISPRi applications in human cells
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Updated: Dec 6, 2025

Protein Extract Preparation and Co-immunoprecipitation from Caenorhabditis elegans
Published on: May 23, 2020
Nader Alerasool1,2, Dmitri Segal1,2, Hunsang Lee1
1Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada.
Abstract:
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.
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