ChIP-Sequencing in Xenopus Embryos

Cold Spring Harb Protoc. 2019 Jan 2;2019(1). doi: 10.1101/pdb.prot097907.

Abstract

Chromatin immunoprecipitation (ChIP) followed by deep sequencing (ChIP-seq) is a powerful technique for mapping in vivo, genome-wide DNA-protein interactions. The interplay between DNA and proteins determines the transcriptional state of the genome. Using specific antibodies for the ChIP, it is possible to generate genome-wide profiles of histone posttranslational modifications, providing insight into the epigenetic memory and developmental potential of cells. The interactions between DNA and proteins involved in epigenetic regulation and transcription are highly dynamic during embryonic development. ChIP-seq allows for a detailed analysis of these dynamic changes in DNA-protein binding during embryogenesis. ChIP-seq is performed on protein epitopes that have been cross-linked to genomic DNA. After shearing the DNA, fragments bound by the (modified) protein of interest are captured with antibodies. The genomic loci of interest are identified by sequencing. Here, we provide a step-by-step ChIP-seq protocol that efficiently captures epitopes from relatively small embryo samples.

MeSH terms

  • Animals
  • Binding Sites
  • Chromatin Immunoprecipitation Sequencing / methods*
  • DNA / chemistry
  • DNA / genetics*
  • DNA / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Gene Expression Regulation, Developmental
  • Protein Binding
  • Xenopus / embryology

Substances

  • DNA-Binding Proteins
  • DNA