Functional interrogation of non-coding DNA through CRISPR genome editing.

Matthew C Canver, Daniel E Bauer, Stuart H Orkin,

Methods (San Diego, Calif.), March 14, 2017

Ologies to interrogate non-coding regions have lagged behind coding regions despite comprising the vast majority of the genome. However, the rapid evolution of clustered regularly interspaced short palindromic repeats (CRISPR)-based genome editing has provided a multitude of novel techniques for laboratory investigation including significant contributions to the toolbox for studying non-coding DNA. CRISPR-mediated loss-of-function strategies rely on direct disruption of the underlying sequence or repression of transcription without modifying the targeted DNA sequence. CRISPR-mediated gain-of-function approaches similarly benefit from methods to alter the targeted sequence through integration of customized sequence into the genome as well as methods to activate transcription. Here we review CRISPR-based loss- and gain-of-function techniques for the interrogation of non-coding DNA.

Copyright © 2017. Published by Elsevier Inc.

Pubmed Link: 28288828

DOI: 10.1016/j.ymeth.2017.03.008