ChIP-chip designs to interrogate the genome of Xenopus embryos for transcription factor binding and epigenetic regulation

PLoS One. 2010 Jan 21;5(1):e8820. doi: 10.1371/journal.pone.0008820.

Abstract

Background: Chromatin immunoprecipitation combined with genome tile path microarrays or deep sequencing can be used to study genome-wide epigenetic profiles and the transcription factor binding repertoire. Although well studied in a variety of cell lines, these genome-wide profiles have so far been little explored in vertebrate embryos.

Principal findings: Here we report on two genome tile path ChIP-chip designs for interrogating the Xenopus tropicalis genome. In particular, a whole-genome microarray design was used to identify active promoters by close proximity to histone H3 lysine 4 trimethylation. A second microarray design features these experimentally derived promoter regions in addition to currently annotated 5' ends of genes. These regions truly represent promoters as shown by binding of TBP, a key transcription initiation factor.

Conclusions: A whole-genome and a promoter tile path microarray design was developed. Both designs can be used to study epigenetic phenomena and transcription factor binding in developing Xenopus embryos.

Publication types

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

MeSH terms

  • Animals
  • Chromatin Immunoprecipitation*
  • Epigenesis, Genetic*
  • Genome*
  • Oligonucleotide Array Sequence Analysis
  • Promoter Regions, Genetic
  • Protein Binding
  • Transcription Factors / metabolism*
  • Xenopus / embryology*
  • Xenopus / genetics

Substances

  • Transcription Factors