In Vivo ChIP-Seq of Nuclear Receptors: A Rough Guide to Transform Frozen Tissues into High-Confidence Genome-Wide Binding Profiles

Methods Mol Biol. 2019:1966:39-70. doi: 10.1007/978-1-4939-9195-2_5.

Abstract

Chromatin immunoprecipitation coupled to next generation sequencing (ChIP-seq) is a powerful tool to map context-dependent genome-wide binding of nuclear hormone receptors and their coregulators. This information can provide important mechanistic insight into where, when and how DNA-protein interactions are linked to target gene regulation. Here we describe a simple, yet reliable ChIP-seq method, including nuclear isolation from frozen tissue samples, cross-linking DNA-protein complexes, chromatin shearing, immunoprecipitation, and purification of ChIP DNA. We also include a standard ChIP-seq data analysis pipeline to elaborate and analyze raw single-end or paired-end sequencing data, including quality control steps, peak calling, annotation, and motif enrichment.

Keywords: ChIP-seq; Chromatin immunoprecipitation; Data analysis; In vivo; Nuclear receptors.

MeSH terms

  • Animals
  • Chromatin Immunoprecipitation / methods*
  • DNA / metabolism
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Sequence Analysis, DNA / methods

Substances

  • Receptors, Cytoplasmic and Nuclear
  • DNA