E-GEOD-43070 - Genome-wide Analysis of Chromatin Interactions in Human Cells

Released on 20 October 2013, last updated on 4 May 2014
Homo sapiens
Protocols (7)
Millions of cis-regulatory sequences have recently been found in the human genome, but the function of most cis-elements are not yet clear, in part due to the difficulty in determining their regulatory targets, which are often located millions of base pairs away and separated by one or more unrelated genes. To address this problem, the Hi-C method has been developed to identify long-range looping interactions in a genome-wide, unbiased fashion. However, current data analysis of Hi-C datasets cannot fully resolve regulatory interactions between enhancers and promoters due to the low resolution. Here, we generated a high-depth Hi-C dataset and applied a new analysis method that offers improved resolution permitting genome-wide identification of nearly one million chromatin interactions. We demonstrated the use of Hi-C to identify target promoters of enhancers regulated by NF-κB signaling and signal-dependent dynamic chromatin interaction at these enhancers in human cells. Surprisingly, our results showed that most NF-κB binding sites are looped to their regulatory targets prior to activation of the signaling pathway, and appear to undergo little change during signaling. This observation suggests that the chromatin organization landscape, once established in a cell type, is rather stable and may influence the selection and activation of target genes by a transcription factor. We performed Hi-C analysis using a human fibroblast cell line IMR90 before and after NF-κB activation. In the meantime, we also performed ChIP-seq experiments to map the location of NF-κB p65 subunit, RNA polymerase II, p300, and several histone modifications (including H3K4me1, H3K4me3, H3K27ac and H3K36me3) in IMR90 cells before and after transient TNF-α stimulation. Additionally, to monitor the dynamic transcription profiles, we also performed Global Run-On sequencing (GRO-seq).
Experiment types
ChIP-seq, other, RNA-seq of coding RNA 
Bing Ren, Fulai Jin, Yan Li
A high-resolution map of the three-dimensional chromatin interactome in human cells. Jin F, Li Y, Dixon JR, Selvaraj S, Ye Z, Lee AY, Yen CA, Schmitt AD, Espinoza CA, Ren B. , Europe PMC 24141950
Exp. designProtocolsVariablesProcessedSeq. reads