A Multiplexed System for Quantitative Comparisons of Chromatin Landscapes

Mol Cell. 2016 Jan 7;61(1):170-80. doi: 10.1016/j.molcel.2015.11.003. Epub 2015 Dec 10.

Abstract

Genome-wide profiling of histone modifications can provide systematic insight into the regulatory elements and programs engaged in a given cell type. However, conventional chromatin immunoprecipitation and sequencing (ChIP-seq) does not capture quantitative information on histone modification levels, requires large amounts of starting material, and involves tedious processing of each individual sample. Here, we address these limitations with a technology that leverages DNA barcoding to profile chromatin quantitatively and in multiplexed format. We concurrently map relative levels of multiple histone modifications across multiple samples, each comprising as few as a thousand cells. We demonstrate the technology by monitoring dynamic changes following inhibition of p300, EZH2, or KDM5, by linking altered epigenetic landscapes to chromatin regulator mutations, and by mapping active and repressive marks in purified human hematopoietic stem cells. Hence, this technology enables quantitative studies of chromatin state dynamics across rare cell types, genotypes, environmental conditions, and drug treatments.

Publication types

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

MeSH terms

  • Chromatin / genetics
  • Chromatin / metabolism*
  • Chromatin Assembly and Disassembly* / drug effects
  • Chromatin Immunoprecipitation / methods*
  • DNA Barcoding, Taxonomic
  • Epigenesis, Genetic / drug effects
  • Gene Expression Profiling
  • Gene Expression Regulation, Leukemic
  • Hematopoietic Stem Cells / metabolism*
  • High-Throughput Nucleotide Sequencing / methods*
  • Histones / genetics
  • Histones / metabolism*
  • Humans
  • K562 Cells
  • Leukemia / genetics
  • Leukemia / metabolism*
  • Multiplex Polymerase Chain Reaction / methods*
  • Mutation

Substances

  • Chromatin
  • Histones