Automated in situ chromatin profiling efficiently resolves cell types and gene regulatory programs

Derek H. Janssens, Steven J. Wu, Jay F. Sarthy, Michael P. Meers, Carrie H. Myers, James M. Olson, Kami Ahmad, Steven Henikoff

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

Background: Our understanding of eukaryotic gene regulation is limited by the complexity of protein-DNA interactions that comprise the chromatin landscape and by inefficient methods for characterizing these interactions. We recently introduced CUT&RUN, an antibody-targeted nuclease cleavage method that profiles DNA-binding proteins, histones and chromatin-modifying proteins in situ with exceptional sensitivity and resolution. Results: Here, we describe an automated CUT&RUN platform and apply it to characterize the chromatin landscapes of human cells. We find that automated CUT&RUN profiles of histone modifications crisply demarcate active and repressed chromatin regions, and we develop a continuous metric to identify cell-type-specific promoter and enhancer activities. We test the ability of automated CUT&RUN to profile frozen tumor samples and find that our method readily distinguishes two pediatric glioma xenografts by their subtype-specific gene expression programs. Conclusions: The easy, cost-effective workflow makes automated CUT&RUN an attractive tool for high-throughput characterization of cell types and patient samples.

Original languageEnglish
Article number74
JournalEpigenetics and Chromatin
Volume11
Issue number1
DOIs
StatePublished - Dec 21 2018

Keywords

  • CUT&RUN
  • Chromatin regulators
  • Histone modifications
  • Transcription factors

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