DSBCapture: in situ capture and sequencing of DNA breaks.
Accepted version
Peer-reviewed
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Repository DOI
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Authors
Lensing, Stefanie V
Marsico, Giovanni
Hänsel-Hertsch, Robert
Lam, Enid Y
Tannahill, David https://orcid.org/0000-0002-3811-6864
Abstract
Double-strand DNA breaks (DSBs) continuously arise and cause mutations and chromosomal rearrangements. Here, we present DSBCapture, a sequencing-based method that captures DSBs in situ and directly maps these at single-nucleotide resolution, enabling the study of DSB origin. DSBCapture shows substantially increased sensitivity and data yield compared with other methods. Using DSBCapture, we uncovered a striking relationship between DSBs and elevated transcription within nucleosome-depleted chromatin.
Description
Keywords
Cell Culture Techniques, Cell Nucleus, Chromatin, DNA, DNA Breaks, Double-Stranded, DNA End-Joining Repair, Epigenesis, Genetic, HeLa Cells, High-Throughput Nucleotide Sequencing, Humans, Keratinocytes, Sensitivity and Specificity, Sequence Analysis, DNA
Journal Title
Nat Methods
Conference Name
Journal ISSN
1548-7091
1548-7105
1548-7105
Volume Title
13
Publisher
Springer Science and Business Media LLC
Publisher DOI
Sponsorship
European Molecular Biology Organization (EMBO) (EMBO ALTF 330-2013)
Cancer Research UK (CB4330)
Cancer Research UK (C14303/A17197)
Cancer Research UK (CB4330)
Cancer Research UK (C14303/A17197)
We thank G. Legube, LBCMCP, Center for Integrative Biology (CBI), Université de Toulouse, Toulouse, France for providing U2OS AID-DIvA cells. We thank the genomic core facility at the Cancer Research UK Cambridge Institute. R.H.-H. acknowledges EMBO for support (EMBO Long-Term Fellowship to R.H.-H.). We acknowledge support from the University of Cambridge and the Cancer Research UK program. The Balasubramanian laboratory is supported by core funding from Cancer Research UK (C14303/A17197 to S.B.) and by an ERC Advanced Grant (S.B.). S.B. is a senior investigator of the Wellcome Trust.