Genome-wide identification of hypoxia-induced enhancer regions

Institute of Molecular Biology, University of Oregon, Eugene, Oregon, United States
DOI
10.7287/peerj.preprints.1393v1
Subject Areas
Biotechnology, Genomics
Keywords
Transcriptional Enhancer, Hypoxia, Next-Generation Sequencing, Massively Parallel Reporter Assay, Method Development
Copyright
© 2015 Kamps-Hughes et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ PrePrints) and either DOI or URL of the article must be cited.
Cite this article
Kamps-Hughes N, Preston JL, Randel MA, Johnson EA. 2015. Genome-wide identification of hypoxia-induced enhancer regions. PeerJ PrePrints 3:e1393v1

Abstract

Here we present a genome-wide method for de novo identification of enhancer regions and apply it to find enhancers that have increased activity after hypoxia. The method links fragmented genomic DNA to the transcription of randomer molecule identifiers and measures the functional enhancer activity of the library by massively parallel sequencing. We transfected a Drosophila melanogaster library into S2 cells in normoxia and hypoxia, and assayed 4,599,881 genomic DNA fragments in parallel. The locations of the enhancer regions strongly correlate with genes up-regulated after hypoxia and previously described enhancers. Novel enhancer regions were identified and integrated with RNAseq data and transcription factor motifs to describe the hypoxic response on a genome-wide basis as a complex regulatory network involving multiple stress-response pathways. This work provides a novel method for high-throughput assay of enhancer activity and the genome-scale identification of hypoxia-activated enhancers in Drosophila.

Author Comment

This is a submission to PeerJ for review.

Supplemental Information

Oligonucleotide sequences used for library construction

DOI: 10.7287/peerj.preprints.1393v1/supp-1

Construct library sequence

DOI: 10.7287/peerj.preprints.1393v1/supp-2

Sequences of identified hypoxia-induced enhancer regions

DOI: 10.7287/peerj.preprints.1393v1/supp-3

Loci of identified baseline enhancer regions

DOI: 10.7287/peerj.preprints.1393v1/supp-4

Computational pipeline for enhancer identification

DOI: 10.7287/peerj.preprints.1393v1/supp-5

RNAseq FPKMs from hypoxic and normoxic S2 cells

DOI: 10.7287/peerj.preprints.1393v1/supp-6