Next-generation sequencing applied to flower development: ChIP-Seq

Methods Mol Biol. 2014:1110:413-29. doi: 10.1007/978-1-4614-9408-9_24.

Abstract

Over the past 20 years, classic genetic approaches have shown that the developmental program underlying flower formation involves a large number of transcriptional regulators. However, the target genes of these transcription factors, as well as the gene regulatory networks they control, remain largely unknown. Chromatin immunoprecipitation coupled to next-generation sequencing (ChIP-Seq), which allows the identification of transcription factor binding sites on a genome-wide scale, has been successfully applied to a number of transcription factors in Arabidopsis. The ChIP-Seq procedure involves chemical cross-linking of proteins to DNA, followed by chromatin fragmentation and immunoprecipitation of specific protein-DNA complexes. The regions of the genome bound by a specific transcription factor can then be identified after next-generation sequencing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / genetics
  • Chromatin Immunoprecipitation / methods*
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification
  • Flowers / cytology
  • Flowers / genetics*
  • Flowers / growth & development*
  • High-Throughput Nucleotide Sequencing / methods*
  • Polymerase Chain Reaction
  • Reproducibility of Results
  • Sequence Analysis, DNA / methods*
  • Tissue Fixation

Substances

  • Chromatin
  • DNA, Plant