A Molecular Chipper technology for CRISPR sgRNA library generation and functional mapping of noncoding regions.

ABSTRACT: Clustered regularly-interspaced palindromic repeats (CRISPR)-based genetic screens using single-guide-RNA (sgRNA) libraries have proven powerful to identify genetic regulators. Applying CRISPR screens to interrogate functional elements in noncoding regions requires generating sgRNA libraries that are densely covering, and ideally inexpensive, easy to implement and flexible for customization. Here we present a Molecular Chipper technology for generating dense sgRNA libraries for genomic regions of interest, and a proof-of-principle screen that identifies novel cis-regulatory domains for miR-142 biogenesis. The Molecular Chipper approach utilizes a combination of random fragmentation and a type III restriction enzyme to derive a densely covering sgRNA library from input DNA. Applying this ap

SUBMITTER: Cheng J 

PROVIDER: S-EPMC4820989 | biostudies-literature | 2016 Mar

REPOSITORIES: biostudies-literature