A high‐throughput ChIP‐Seq for large‐scale chromatin studies

Molecular Systems Biology

Christophe D Chabbert¹  Sophie H Adjalley¹  Bernd Klaus¹  Emilie S Fritsch¹  Ishaan Gupta¹  Vicent Pelechano¹ 

[1] European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany

关键词: ChIP‐Seq;    chromatin;    high‐throughput;    histone marks;    histone methyltransferase;   

DOI  :  10.15252/msb.20145776

来源: Wiley

【 摘 要 】

Abstract

We present a modified approach of chromatin immuno-precipitation followed by sequencing (ChIP-Seq), which relies on the direct ligation of molecular barcodes to chromatin fragments, thereby permitting experimental scale-up. With Bar-ChIP now enabling the concurrent profiling of multiple DNA–protein interactions, we report the simultaneous generation of 90 ChIP-Seq datasets without any robotic instrumentation. We demonstrate that application of Bar-ChIP to a panel of Saccharomyces cerevisiae chromatin-associated mutants provides a rapid and accurate genome-wide overview of their chromatin status. Additionally, we validate the utility of this technology to derive novel biological insights by identifying a role for the Rpd3S complex in maintaining H3K14 hypo-acetylation in gene bodies. We also report an association between the presence of intragenic H3K4 tri-methylation and the emergence of cryptic transcription in a Set2 mutant. Finally, we uncover a crosstalk between H3K14 acetylation and H3K4 methylation in this mutant. These results show that Bar-ChIP enables biological discovery through rapid chromatin profiling at single-nucleosome resolution for various conditions and protein modifications at once.

Synopsis

A new approach provides a rapid and accurate genome-wide overview of the chromatin status of multiple yeast chromatin-associated mutants at once. The simultaneous profiling of epigenetic marks in the mutants is achieved by multiplex immuno-precipitation of barcoded chromatin samples.

• Bar-ChIP is based on the immuno-precipitation of barcoded chromatin and permits sample multiplexing, thereby increasing the throughput of ChIP-Seq experiments.
• Application of the method to yeast chromatin-associated mutants enabled the concurrent generation of 90 ChIP-Seq datasets without the need for robotic instrumentation.
• The rapid chromatin profiling of the mutants at single-nucleosome resolution uncovered an association between intragenic H3K4 tri-methylation and cryptic transcription in set2∆.

【 授权许可】

CC BY   
© 2015 The Authors. Published under the terms of the CC BY 4.0 license

Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.