Capture of associated targets on chromatin links long-distance chromatin looping to transcriptional coordination

Nat Commun. 2016 Sep 16:7:12893. doi: 10.1038/ncomms12893.

Abstract

Here we describe a sensitive and novel method of identifying endogenous DNA-DNA interactions. Capture of Associated Targets on CHromatin (CATCH) uses efficient capture and enrichment of specific genomic loci of interest through hybridization and subsequent purification via complementary biotinylated oligonucleotide. The CATCH assay requires no enzymatic digestion or ligation, requires little starting material, provides high-quality data, has excellent reproducibility and is completed in less than 24 h. Efficacy is demonstrated through capture of three disparate loci, which demonstrate unique subsets of long-distance chromatin interactions enriched for both enhancer marks and oestrogen receptor-binding sites. In each experiment, CATCH-seq peaks representing long-distance chromatin interactions were centred near the TSS of genes, and, critically, the genes identified as physically interacting are shown to be transcriptionally coexpressed. These interactions could potentially create transcriptional hubs for the regulation of gene expression programmes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Chromatin Immunoprecipitation
  • Chromatin*
  • DNA
  • Endoplasmic Reticulum
  • Eukaryotic Initiation Factor-4A / genetics
  • Eukaryotic Initiation Factor-4A / metabolism
  • Gene Expression Regulation
  • Genomics
  • Humans
  • Promoter Regions, Genetic
  • Protein Binding
  • Sequence Analysis, DNA / methods

Substances

  • Chromatin
  • DNA
  • Eukaryotic Initiation Factor-4A