PT  - JOURNAL ARTICLE
AU  - Ali Shariati
AU  - Antonia Dominguez
AU  - Marius Wernig
AU  - Lei S. Qi
AU  - Jan M. Skotheim
TI  - Reversible inhibition of specific transcription factor-DNA interactions using CRISPR
AID  - 10.1101/282681
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 282681
4099  - http://biorxiv.org/content/early/2018/03/15/282681.short
4100  - http://biorxiv.org/content/early/2018/03/15/282681.full
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.