CRISPR Inversion of CTCF Sites Alters Genome Topology and Enhancer/Promoter Function.

ABSTRACT: CTCF and the associated cohesin complex play a central role in insulator function and higher-order chromatin organization of mammalian genomes. Recent studies identified a correlation between the orientation of CTCF-binding sites (CBSs) and chromatin loops. To test the functional significance of this observation, we combined CRISPR/Cas9-based genomic-DNA-fragment editing with chromosome-conformation-capture experiments to show that the location and relative orientations of CBSs determine the specificity of long-range chromatin looping in mammalian genomes, using protocadherin (Pcdh) and β-globin as model genes. Inversion of CBS elements within the Pcdh enhancer reconfigures the topology of chromatin loops between the distal enhancer and target promoters and alters gene-expression patterns.

SUBMITTER: Guo Y 

PROVIDER: S-EPMC4642453 | biostudies-literature | 2015 Aug

REPOSITORIES: biostudies-literature