Abstract

BackgroundChromatin conformation capture with high-throughput sequencing (Hi-C) is a technique that measures the in vivo intensity of interactions between all pairs of loci in the genome. Most conventional analyses of Hi-C data focus on the detection of statistically significant interactions. However, an alternative strategy involves identifying significant changes in the interaction intensity (i.e., differential interactions) between two or more biological conditions. This is more statistically rigorous and may provide more biologically relevant results.ResultsHere, we present the diffHic software package for the detection of differential interactions from Hi-C data. diffHic provides methods for read pair alignment and processing, counting into bin pairs, filtering out low-abundance events and normalization of trended or CNV-driven biases. It uses the statistical framework of the edgeR package to model biological variability and to test for significant differences between conditions. Several options for the visualization of results are also included. The use of diffHic is demonstrated with real Hi-C data sets. Performance against existing methods is also evaluated with simulated data.ConclusionsOn real data, diffHic is able to successfully detect interactions with significant differences in intensity between biological conditions. It also compares favourably to existing software tools on simulated data sets. These results suggest that diffHic is a viable approach for differential analyses of Hi-C data.Electronic supplementary materialThe online version of this article (doi:10.1186/s12859-015-0683-0) contains supplementary material, which is available to authorized users.

Highlights

  • Chromatin conformation capture with high-throughput sequencing (Hi-C) is a technique that measures the in vivo intensity of interactions between all pairs of loci in the genome

  • Introduction to the real data sets The diffHic pipeline can be applied on any Hi-C data set containing biological replicates across multiple conditions, where the aim is to detect DIs between conditions

  • The use of diffHic will be demonstrated on three Hi-C data sets

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Summary

Introduction

Chromatin conformation capture with high-throughput sequencing (Hi-C) is a technique that measures the in vivo intensity of interactions between all pairs of loci in the genome. An alternative strategy involves identifying significant changes in the interaction intensity (i.e., differential interactions) between two or more biological conditions. This is more statistically rigorous and may provide more biologically relevant results. Chromatin conformation capture with high-throughput sequencing (Hi-C) is a technique that is widely used to study global chromatin organization in vivo [1]. Each complex may contain multiple restriction fragments, corresponding to an interaction between the associated genomic loci. Proximity ligation is performed between the ends of the restriction fragments.

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