A high-throughput ChIP-Seq for large-scale chromatin studies.

Autor: Chabbert CD; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany., Adjalley SH; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany., Klaus B; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany., Fritsch ES; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany., Gupta I; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany., Pelechano V; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany pelechan@embl.de larsms@embl.de., Steinmetz LM; European Molecular Biology Laboratory, Genome Biology Unit, Heidelberg, Germany Stanford Genome Technology Center, Palo Alto, CA, USA Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA pelechan@embl.de larsms@embl.de.
Jazyk: angličtina
Zdroj: Molecular systems biology [Mol Syst Biol] 2015 Jan 12; Vol. 11 (1), pp. 777. Date of Electronic Publication: 2015 Jan 12.
DOI: 10.15252/msb.20145776
Abstrakt: 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.
(© 2015 The Authors. Published under the terms of the CC BY 4.0 license.)
Databáze: MEDLINE
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