Hi-C on the OCIAML-2 Cell Line
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ABSTRACT:
PROVIDER: EGAS00001004743 | EGA |
REPOSITORIES: EGA
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Project description:Capture Hi-C (CHi-C) is a state-of-the art method for profiling chromosomal interactions involving targeted regions of interest (such as gene promoters) globally and at high resolution. Signal detection in CHi-C data involves a number of statistical challenges that are not observed when using other Hi-C-like techniques. We present a background model, and algorithms for normalisation and multiple testing that are specifically adapted to CHi-C experiments, in which many spatially dispersed regions are captured, such as in Promoter CHi-C. We implement these procedures in CHiCAGO (http://regulatorygenomicsgroup.org/chicago), an open-source package for robust interaction detection in CHi-C. We validate CHiCAGO by showing that promoter-interacting regions detected with this method are enriched for regulatory features and disease-associated SNPs. Three human CHi-C biological replicates were generated (comprising 1, 2and 3 technical replicates). Two mouse CHi-C biological replicates were generated (both comprising three technical replicates) and a mouse Hi-C dataset. The publicly available HiCUP pipeline (doi: 10.12688/f1000research.7334.1) was used to process the raw sequencing reads. This pipeline was used to map the read pairs against the mouse (mm9) and human (hg19) genomes, to filter experimental artefacts (such as circularized reads and re-ligations), and to remove duplicate reads. For the CHi-C data, the resulting BAM files were processed into CHiCAGO input files, retaining only those read pairs that mapped, at least on one end, to a captured bait. CHiCAGO then identified Hi-C restriction fragments interacting, with statistical significant, to captured baits.
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Project description:Perturbation of domain boundaries causes restructuration of domains and gene expression misregulation. Structural proteins such as CTCF and the cohesin complex have been identified enriched at domain boundaries. However, we still lack a complete understanding of the function of boundaries and what makes one boundary more effective at insulating genomic interactions than others. To understand how domains are structured, we experimentally classified boundaries according to their accessibility degree as a proxy of protein occupancy in K562 human cells. To determine if accessibility indeed reflects more robust boundaries and domains, we experimentally explored whether domain boundaries exhibit different degrees of accessibility in K562 cell line. To do so, we performed Hi-C experiments varying the incubation time of digestion with the restriction enzyme. The incorporation of this digestion time course into the Hi-C enables us to directly assess boundary accessibility and TAD formation from the same experiments.
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