DSBCapture: in situ capture and sequencing of DNA breaks.
Lensing, Stefanie V
Lam, Enid Y
Nature Publishing Group
MetadataShow full item record
Lensing, S. V., Marsico, G., Hänsel-Hertsch, R., Lam, E. Y., Tannahill, D., & Balasubramanian, S. (2016). DSBCapture: in situ capture and sequencing of DNA breaks.. Nature methods, 13 855-857. https://doi.org/10.1038/nmeth.3960
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.
Hela Cells, Cell Nucleus, Chromatin, Keratinocytes, Humans, DNA, Cell Culture Techniques, Sensitivity and Specificity, Sequence Analysis, DNA, Epigenesis, Genetic, DNA Breaks, Double-Stranded, High-Throughput Nucleotide Sequencing, DNA End-Joining Repair
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.
European Molecular Biology Organization (EMBO) (EMBO ALTF 330-2013)
Cancer Research UK (CB4330)
Cancer Research UK (C14303_do not transfer)
External DOI: https://doi.org/10.1038/nmeth.3960
This record's URL: https://www.repository.cam.ac.uk/handle/1810/259980