ChIA-PET analysis of transcriptional chromatin interactions

Methods. 2012 Nov;58(3):289-99. doi: 10.1016/j.ymeth.2012.08.009. Epub 2012 Aug 25.

Abstract

Long-range chromatin contacts between specific DNA regulatory elements play a pivotal role in gene expression regulation, and a global characterization of these interactions in the 3-dimensional (3D) chromatin structure is imperative in understanding signaling networks and cell states. Chromatin Interaction Analysis using Paired-End Tag sequencing (ChIA-PET) is a method which converts functional chromatin structure into millions of short tag sequences. Combining Chromatin Immunoprecipitation (ChIP), proximity ligation and high-throughput sequencing, ChIA-PET provides a global and unbiased interrogation of higher-order chromatin structures associated with specific protein factors. Here, we describe the detailed procedures of the ChIA-PET methodology, unraveling transcription-associated chromatin contacts in a model human cell line.

Publication types

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

MeSH terms

  • Base Sequence
  • Cell Line
  • Chromatin / genetics*
  • Chromatin / ultrastructure
  • Chromatin Immunoprecipitation
  • Chromosome Mapping
  • Cross-Linking Reagents / chemistry
  • DNA / chemistry
  • DNA / genetics
  • DNA / isolation & purification
  • Epistasis, Genetic
  • Gene Library
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Sequence Analysis, DNA*
  • Sonication
  • Succinimides / chemistry

Substances

  • Chromatin
  • Cross-Linking Reagents
  • Succinimides
  • ethylene glycolylbis(succinimidyl succinate)
  • DNA