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qDSB-Seq is a general method for genome-wide quantification of DNA double-strand breaks using sequencing

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dc.abstract.enDNA double-strand breaks (DSBs) are among the most lethal types of DNA damage and frequently cause genome instability. Sequencing-based methods for mapping DSBs have been developed but they allow measurement only of relative frequencies of DSBs between loci, which limits our understanding of the physiological relevance of detected DSBs. Here we propose quantitative DSB sequencing (qDSB-Seq), a method providing both DSB frequencies per cell and their precise genomic coordinates. We induce spike-in DSBs by a site-specific endonuclease and use them to quantify detected DSBs (labeled, e.g., using i-BLESS). Utilizing qDSB-Seq, we determine numbers of DSBs induced by a radiomimetic drug and replication stress, and reveal two orders of magnitude differences in DSB frequencies. We also measure absolute frequencies of Top1-dependent DSBs at natural replication fork barriers. qDSB-Seq is compatible with various DSB labeling methods in different organisms and allows accurate comparisons of absolute DSB frequencies across samples.
dc.affiliationUniwersytet Warszawski
dc.contributor.authorRowicka, Maga
dc.contributor.authorNde, Jules
dc.contributor.authorFongang, Bernard
dc.contributor.authorPasero, Philippe
dc.contributor.authorZhu, Yingjie
dc.contributor.authorYousefi, Razie
dc.contributor.authorPardo, Benjamin
dc.contributor.authorForey, Romain
dc.contributor.authorSkrzypczak, Magdalena
dc.contributor.authorBiernacka, Anna
dc.contributor.authorDojer, Norbert
dc.contributor.authorGinalski, Krzysztof
dc.date.accessioned2024-01-25T18:42:59Z
dc.date.available2024-01-25T18:42:59Z
dc.date.issued2019
dc.description.financeNie dotyczy
dc.description.volume10
dc.identifier.doi10.1038/S41467-019-10332-8
dc.identifier.issn2041-1723
dc.identifier.urihttps://repozytorium.uw.edu.pl//handle/item/117641
dc.identifier.weblinkhttp://dx.doi.org/10.1038/s41467-019-10332-8
dc.languageeng
dc.pbn.affiliationcomputer and information sciences
dc.relation.ispartofNature Communications
dc.relation.pages2313:1-2313:11
dc.rightsClosedAccess
dc.sciencecloudnosend
dc.subject.enDouble-strand DNA breaks
dc.subject.enGenomic instability
dc.subject.enNext-generation sequencing
dc.subject.enStalled forks
dc.titleqDSB-Seq is a general method for genome-wide quantification of DNA double-strand breaks using sequencing
dc.typeJournalArticle
dspace.entity.typePublication