A Comprehensive Toolbox to Analyze Enhancer-Promoter Functions

Methods Mol Biol. 2021:2351:3-22. doi: 10.1007/978-1-0716-1597-3_1.

Abstract

Knowledge in gene transcription and chromatin regulation has been intensely studied for decades, but thanks to next-generation sequencing (NGS) techniques there has been a major leap forward in the last few years. Historically, identification of specific enhancer elements has led to the identification of master transcription factors (TFs) in the 1990s. Genetic and biochemical experiments have identified the key regulators controlling RNA polymerase II (RNAPII) transcription and structurally analyses have elucidated detailed mechanisms. NGS and the development of chromatin immunoprecipitation (ChIP) have accelerated the gain of knowledge in the recent years. By now, we have a dazzling wealth of techniques that are currently used to put gene expression into a genome-wide context. This book is an attempt to assemble useful protocols for many researchers within and nearby research areas. In general, these innovative techniques focus on enhancer and promoter studies. The techniques should also be of interest for related fields such as DNA repair and replication.

Keywords: DNA methylation; Enhancers; Histone variants; Promoters; Transcription; eRNAs.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Animals
  • Binding Sites
  • Chromatin / genetics
  • Chromatin / metabolism
  • Chromatin Immunoprecipitation
  • DNA Methylation
  • Enhancer Elements, Genetic*
  • Epigenesis, Genetic
  • Gene Expression Regulation*
  • Histones / metabolism
  • Humans
  • Promoter Regions, Genetic*
  • Protein Binding
  • RNA Polymerase II / metabolism
  • Transcription, Genetic*

Substances

  • Chromatin
  • Histones
  • RNA Polymerase II