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Nature. 2019 Feb;566(7745):558-562. doi: 10.1038/s41586-019-0949-1. Epub 2019 Feb 18.

Multiplex chromatin interactions with single-molecule precision.

Author information

1
The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA.
2
Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, CT, USA.
3
School of Optometry and Ophthalmology, Wenzhou Medical University, Wenzhou, China.
4
The Jackson Laboratory for Genomic Medicine, Farmington, CT, USA. yijun.ruan@jax.org.
5
Department of Genetics and Genome Sciences, University of Connecticut Health Center, Farmington, CT, USA. yijun.ruan@jax.org.
6
Huazhong Agricultural University, Wuhan, China. yijun.ruan@jax.org.

Abstract

The genomes of multicellular organisms are extensively folded into 3D chromosome territories within the nucleus1. Advanced 3D genome-mapping methods that combine proximity ligation and high-throughput sequencing (such as chromosome conformation capture, Hi-C)2, and chromatin immunoprecipitation techniques (such as chromatin interaction analysis by paired-end tag sequencing, ChIA-PET)3, have revealed topologically associating domains4 with frequent chromatin contacts, and have identified chromatin loops mediated by specific protein factors for insulation and regulation of transcription5-7. However, these methods rely on pairwise proximity ligation and reflect population-level views, and thus cannot reveal the detailed nature of chromatin interactions. Although single-cell Hi-C8 potentially overcomes this issue, this method may be limited by the sparsity of data that is inherent to current single-cell assays. Recent advances in microfluidics have opened opportunities for droplet-based genomic analysis9 but this approach has not yet been adapted for chromatin interaction analysis. Here we describe a strategy for multiplex chromatin-interaction analysis via droplet-based and barcode-linked sequencing, which we name ChIA-Drop. We demonstrate the robustness of ChIA-Drop in capturing complex chromatin interactions with single-molecule precision, which has not been possible using methods based on population-level pairwise contacts. By applying ChIA-Drop to Drosophila cells, we show that chromatin topological structures predominantly consist of multiplex chromatin interactions with high heterogeneity; ChIA-Drop also reveals promoter-centred multivalent interactions, which provide topological insights into transcription.

PMID:
30778195
DOI:
10.1038/s41586-019-0949-1

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