ChIP-enriched in silico targets (ChEST), a ChIP-on-chip approach applied to analyzing skeletal muscle genes

Methods Mol Biol. 2012:798:543-53. doi: 10.1007/978-1-61779-343-1_33.

Abstract

Mapping the cis-regulatory modules (CRMs) to which bind myogenic transcription factors is an -obligatory step towards understanding gene regulatory networks governing muscle development and function. This can be achieved in silico or by chromatin immunoprecipitation (ChIP) approaches. We have developed a ChIP-enriched in silico targets (ChEST) strategy designed for mapping the CRMs by combining in silico and ChIP methods. ChEST involves a software-assisted prediction of transcription factor (TF) - specific CRMs, which are spotted to produce a computed genomic CRM microarray. In parallel, the in vivo pool of targets of a given TF is isolated by ChIP and used as a probe for hybridization with the array generated. Here we describe ChEST strategy applied to identify direct targets of Myogenic Enhancer Factor, Dmef2 in Drosophila embryos.

MeSH terms

  • Animals
  • Chromatin Immunoprecipitation / methods*
  • Computational Biology
  • DNA Probes / chemical synthesis
  • Drosophila / genetics
  • Embryo, Nonmammalian / metabolism
  • Enhancer Elements, Genetic
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Developmental
  • Muscle, Skeletal / metabolism*
  • Nucleic Acid Amplification Techniques
  • Nucleic Acid Hybridization / methods
  • Oligonucleotide Array Sequence Analysis / methods*
  • Regulatory Sequences, Nucleic Acid / genetics*

Substances

  • DNA Probes