Crosetto, NicolaMitra, AbhishekSilva, Maria JoaoBienko, MagdaDojer, NorbertWang, QiKaraca, ElifChiarle, RobertoSkrzypczak, MagdalenaGinalski, KrzysztofPasero, PhilippeRowicka, MagaDikic, Ivan2014-03-102013Crosetto, N., A. Mitra, M. J. Silva, M. Bienko, N. Dojer, Q. Wang, E. Karaca, et al. 2013. “Nucleotide-resolution DNA double-strand breaks mapping by next-generation sequencing.” Nature methods 10 (4): 361-365. doi:10.1038/nmeth.2408. http://dx.doi.org/10.1038/nmeth.2408.1548-7091http://nrs.harvard.edu/urn-3:HUL.InstRepos:11878917We present a genome-wide method to map DNA double-strand breaks (DSBs) at nucleotide resolution by direct in situ breaks labeling, enrichment on streptavidin, and next-generation sequencing (BLESS). We comprehensively validated and tested BLESS using different human and mouse cells, DSBs-inducing agents, and sequencing platforms. BLESS was able to detect telomere ends, Sce endonuclease-induced DSBs, and complex genome-wide DSBs landscapes. As a proof of principle, we characterized the genomic landscape of sensitivity to replication stress in human cells, and identified over two thousand non-uniformly distributed aphidicolin-sensitive regions (ASRs) overrepresented in genes and enriched in satellite repeats. ASRs were also enriched in regions rearranged in human cancers, with many cancer-associated genes exhibiting high sensitivity to replication stress. Our method is suitable for genome-wide mapping of DSBs in various cells and experimental conditions with a specificity and resolution unachievable by current techniques.en-USJournal Article2014-03-1010.1038/nmeth.2408