The Study of Protein-DNA Interactions in CD4+ T-Cells Using ChIPmentation

Methods Mol Biol. 2021:2285:201-216. doi: 10.1007/978-1-0716-1311-5_17.

Abstract

Chromatin immunoprecipitation (ChIP) coupled with high-throughput sequencing (ChIP-seq) is an invaluable method to profile of enrichment of histone modifications and transcription factor binding sites across the genome. However, standard ChIP-seq protocols require large numbers of cells (>107) as starting material, which are often impossible to obtain for rare immune populations. Here we describe a streamlined ChIP protocol optimised for small cell numbers in conjunction with transposon-tagging mediated sequencing library preparation (ChIPmentation) which allows the analysis of samples of as low as 105 cells.

Keywords: ChIP; ChIPmentation; Chromatin immunoprecipitation; Transcription factor.

MeSH terms

  • Animals
  • Binding Sites
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism*
  • Cells, Cultured
  • Chromatin Immunoprecipitation*
  • DNA / metabolism*
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Protein Binding
  • Research Design
  • Transcription Factors / metabolism*
  • Workflow

Substances

  • Transcription Factors
  • DNA