RT Journal Article
SR Electronic
T1 Reversible inhibition of specific transcription factor-DNA interactions using CRISPR
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 282681
DO 10.1101/282681
A1 Ali Shariati
A1 Antonia Dominguez
A1 Marius Wernig
A1 Lei S. Qi
A1 Jan M. Skotheim
YR 2018
UL http://biorxiv.org/content/early/2018/03/15/282681.abstract
AB The control of gene expression by transcription factor binding sites frequently determines phenotype. However, it has been difficult to assay the function of single transcription factor binding sites within larger transcription networks. Here, we developed such a method by using deactivated Cas9 to disrupt binding to specific sites on the genome. Since CRISPR guide RNAs are longer than transcription factor binding sites, flanking sequence can be used to target specific sites. Targeting deactivated Cas9 to a specific Oct4 binding site in the Nanog promoter blocked Oct4 binding, reduced Nanog expression, and slowed division. Multiple guide RNAs allows simultaneous inhibition of multiple binding sites and conditionally-destabilized dCas9 allows rapid reversibility. The method is a novel high-throughput approach to systematically interrogate cis-regulatory function within complex regulatory networks.