Chromatin immunoprecipitation assay to identify genomic binding sites of regulatory factors

Meike Wagner, Johannes Jung, Michael Koslowski, Özlem Türeci, Vijay K. Tiwari, Ugur Sahin

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

DNA–protein interactions are vital to fundamental cellular events including transcription, replication, DNA repair, and recombination. Thus, their study holds the key to our understanding of mechanisms underlying normal development and homeostasis as well as disease. Transcriptional regulation is a highly complex pro- cess that involves recruitment of numerous factors resulting in formation of multi-protein complexes at gene promoters to regulate gene expression. The studied proteins can be, for example, transcription factors, epigenetic regulators, co-activators, co-repressors, or ligand-activated nuclear receptors as estrogen receptor-α (ERα) bound either directly to the DNA or indirectly by interaction with other DNA-bound factors. Chromatin immunoprecipitation (ChIP) assay is a powerful method to study interactions of pro- teins and a specifi c genomic DNA region. Recruitment of ERα to promoters of estrogen- dependent genes is a common mechanism to activate or enhance gene transcription in breast cancer thus promoting tumor progression. In this chapter, we demonstrate a stepwise protocol for ChIP assay using binding of ERα to its genomic targets after stimulation with 17β-estradiol (E 2 ) in breast cancer cells as an example.
Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.,U.S.
Pages53-65
Number of pages13
DOIs
Publication statusPublished - 01 Jan 2016

Publication series

NameMethods in Molecular Biology
Volume1366

Keywords

  • Cancer
  • ChIP assay
  • Chromatin
  • DNA-protein interactions
  • Estrogenreceptorα
  • Estrogensignaling
  • Gene regulation

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