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

Epigenetics & Chromatin

Steven J. Wu¹  Michael P. Meers¹  Jay F. Sarthy¹  Steven Henikoff¹  Kami Ahmad¹  Derek H. Janssens¹  James M. Olson²  Carrie H. Myers³ 

[1] Basic Sciences Division, Fred Hutchinson Cancer Research Center;Cancer and Blood Disorder Center, Seattle Children’s Hospital;Clinical Research Division, Fred Hutchinson Cancer Research Center;

关键词: CUT&RUN;    Histone modifications;    Transcription factors;    Chromatin regulators;   

DOI  :  10.1186/s13072-018-0243-8

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown