CRISPR Interference Efficiently Induces Specific and Reversible Gene Silencing in Human IPSCs

Overview

Journal Cell Stem Cell

Publisher Cell Press

Specialty Cell Biology

Date 2016 Mar 15

PMID 26971820

Citations 258

Authors

Mohammad A Mandegar

Nathaniel Huebsch

Ekaterina B Frolov

Edward Shin

Annie Truong

Michael P Olvera

Amanda H Chan

Yuichiro Miyaoka

Kristin Holmes

C Ian Spencer

Luke M Judge

David E Gordon

Tilde V Eskildsen

Jacqueline E Villalta

Max A Horlbeck

Luke A Gilbert

Nevan J Krogan

Soren P Sheikh

Jonathan S Weissman

Lei S Qi

Po-Lin So

Bruce R Conklin

Affiliations

Soon will be listed here.

Abstract

Developing technologies for efficient and scalable disruption of gene expression will provide powerful tools for studying gene function, developmental pathways, and disease mechanisms. Here, we develop clustered regularly interspaced short palindromic repeat interference (CRISPRi) to repress gene expression in human induced pluripotent stem cells (iPSCs). CRISPRi, in which a doxycycline-inducible deactivated Cas9 is fused to a KRAB repression domain, can specifically and reversibly inhibit gene expression in iPSCs and iPSC-derived cardiac progenitors, cardiomyocytes, and T lymphocytes. This gene repression system is tunable and has the potential to silence single alleles. Compared with CRISPR nuclease (CRISPRn), CRISPRi gene repression is more efficient and homogenous across cell populations. The CRISPRi system in iPSCs provides a powerful platform to perform genome-scale screens in a wide range of iPSC-derived cell types, dissect developmental pathways, and model disease.

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