ChIP-DIP maps binding of hundreds of proteins to DNA simultaneously and identifies diverse gene regulatory elements

Nat Genet. 2024 Dec;56(12):2827-2841. doi: 10.1038/s41588-024-02000-5. Epub 2024 Nov 25.

Abstract

Gene expression is controlled by dynamic localization of thousands of regulatory proteins to precise genomic regions. Understanding this cell type-specific process has been a longstanding goal yet remains challenging because DNA-protein mapping methods generally study one protein at a time. Here, to address this, we developed chromatin immunoprecipitation done in parallel (ChIP-DIP) to generate genome-wide maps of hundreds of diverse regulatory proteins in a single experiment. ChIP-DIP produces highly accurate maps within large pools (>160 proteins) for all classes of DNA-associated proteins, including modified histones, chromatin regulators and transcription factors and across multiple conditions simultaneously. First, we used ChIP-DIP to measure temporal chromatin dynamics in primary dendritic cells following LPS stimulation. Next, we explored quantitative combinations of histone modifications that define distinct classes of regulatory elements and characterized their functional activity in human and mouse cell lines. Overall, ChIP-DIP generates context-specific protein localization maps at consortium scale within any molecular biology laboratory and experimental system.

MeSH terms

  • Animals
  • Binding Sites
  • Cell Line
  • Chromatin Immunoprecipitation* / methods
  • Chromatin* / genetics
  • Chromatin* / metabolism
  • DNA* / genetics
  • DNA* / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Dendritic Cells / metabolism
  • Gene Expression Regulation
  • Histones / genetics
  • Histones / metabolism
  • Humans
  • Mice
  • Protein Binding
  • Regulatory Sequences, Nucleic Acid
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Chromatin
  • DNA
  • Histones
  • Transcription Factors
  • DNA-Binding Proteins