Chromatin immunoprecipitation in early mouse embryos

Estela G. García-González, Bladimir Roque-Ramirez, Carlos Palma-Flores, J. Manuel Hernández-Hernández

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Epigenetic regulation is achieved at many levels by different factors such as tissue-specific transcription factors, members of the basal transcriptional apparatus, chromatin-binding proteins, and noncoding RNAs. Importantly, chromatin structure dictates the availability of a specific genomic locus for transcriptional activation as well as the efficiency with which transcription can occur. Chromatin immunoprecipitation (ChIP) is a method that allows elucidating gene regulation at the molecular level by assessing if chromatin modifications or proteins are present at a specific locus. Initially, the majority of ChIP experiments were performed on cultured cell lines and more recently this technique has been adapted to a variety of tissues in different model organisms. Using ChIP on mouse embryos, it is possible to document the presence or absence of specific proteins and chromatin modifications at genomic loci in vivo during mammalian development and to get biological meaning from observations made on tissue culture analyses. We describe here a ChIP protocol on freshly isolated mouse embryonic somites for in vivo analysis of muscle specific transcription factor binding on chromatin. This protocol has been easily adapted to other mouse embryonic tissues and has also been successfully scaled up to perform ChIP-Seq.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages145-155
Number of pages11
DOIs
StatePublished - 2018

Publication series

NameMethods in Molecular Biology
Volume1752
ISSN (Print)1064-3745

Keywords

  • Chromatin immunoprecipitation
  • Embryo
  • Epigenetics
  • Gene regulation
  • MyoD
  • Myogenesis
  • Myogenin
  • Somites

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