DSBCapture: in situ capture and sequencing of DNA breaks

Abstract

Double-strand DNA breaks capture (DSBCapture) identifies in situ DSBs via the ligation of an Illumina adaptor into the break site and shows no bias for chromatin state or base composition. A genome-wide DSB profile shows breaks occurring more frequently in euchromatin and transcriptionally active regions. 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.