In situ functional dissection of RNA cis-regulatory elements by multiplex CRISPR-Cas9 genome engineering.

ABSTRACT: RNA regulatory elements (RREs) are an important yet relatively under-explored facet of gene regulation. Deciphering the prevalence and functional impact of this post-transcriptional control layer requires technologies for disrupting RREs without perturbing cellular homeostasis. Here we describe genome-engineering based evaluation of RNA regulatory element activity (GenERA), a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 platform for in situ high-content functional analysis of RREs. We use GenERA to survey the entire regulatory landscape of a 3'UTR, and apply it in a multiplex fashion to analyse combinatorial interactions between sets of miRNA response elements (MREs), providing strong evidence for cooperative activity. We also employ this technology to probe the

SUBMITTER: Wu Q 

PROVIDER: S-EPMC5727397 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature