A chromatin integration labelling method enables epigenomic profiling with lower input

Nat Cell Biol. 2019 Feb;21(2):287-296. doi: 10.1038/s41556-018-0248-3. Epub 2018 Dec 10.

Abstract

Chromatin plays a crucial role in gene regulation, and chromatin immunoprecipitation followed by sequencing (ChIP-seq) has been the standard technique for examining protein-DNA interactions across the whole genome. However, it is difficult to obtain epigenomic information from limited numbers of cells by ChIP-seq because of sample loss during chromatin preparation and inefficient immunoprecipitation. In this study, we established an immunoprecipitation-free epigenomic profiling method named chromatin integration labelling (ChIL), which enables the amplification of genomic sequences closely associated with the target molecules before cell lysis. Using ChIL followed by sequencing (ChIL-seq), we reliably detected the distributions of histone modifications and DNA-binding factors in 100-1,000 cells. In addition, ChIL-seq successfully detected genomic regions associated with histone marks at the single-cell level. Thus, ChIL-seq offers an alternative method to ChIP-seq for epigenomic profiling using small numbers of cells, in particular, those attached to culture plates and after immunofluorescence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cell Line, Tumor
  • Chromatin / genetics*
  • Chromatin / metabolism
  • DNA Probes / genetics*
  • DNA Probes / metabolism
  • Epigenomics / methods*
  • Gene Expression Profiling / methods*
  • Histones / metabolism
  • Humans
  • Lysine / metabolism
  • MCF-7 Cells
  • Methylation
  • Mice
  • Myoblasts / cytology
  • Myoblasts / metabolism*
  • Reproducibility of Results
  • Sequence Analysis, DNA / methods*
  • Single-Cell Analysis / methods

Substances

  • Chromatin
  • DNA Probes
  • Histones
  • Lysine