期刊
EPIGENETICS & CHROMATIN
卷 11, 期 -, 页码 -出版社
BMC
DOI: 10.1186/s13072-018-0243-8
关键词
CUT&RUN; Histone modifications; Transcription factors; Chromatin regulators
资金
- Chan-Zuckerberg Initiative
- Howard Hughes Medical Institute
- Damon Runyon-Sohn Foundation Fellowship
BackgroundOur 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.ResultsHere, 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.ConclusionsThe easy, cost-effective workflow makes automated CUT&RUN an attractive tool for high-throughput characterization of cell types and patient samples.
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