Genomewide nucleotide-resolution mapping of single-strand breaks and lesions by GLOE-Seq [digestion]

Numerous methods are available for the mapping of DNA lesions ranging from double-strand breaks (DSBs) to incorporated ribonucleotides. We now present a technology based on the Genomewide Ligation of 3’-OH Ends (GLOE-Seq) and an associated computational pipeline designed for single-stranded breaks (SSBs), but versatile enough to be applied to any lesion that is convertible into a free 3’-OH terminus. We demonstrate its applicability to the mapping of Okazaki fragments without prior size selection and detect biases and asymmetries in the distribution of spontaneous SSBs in budding yeast and human DNA.目前已有多种方法可用于绘制从双链断裂（double-strand breaks, DSBs）到掺入核糖核苷酸的各类DNA损伤图谱。本文提出一种基于3’-OH端全基因组连接法（Genomewide Ligation of 3’-OH Ends, GLOE-Seq）的技术，以及配套的计算流程：该技术最初设计用于单链断裂（single-stranded breaks, SSBs）的检测，但通用性极强，可应用于所有可转化为游离3’-OH末端的损伤类型。我们验证了该技术在无需预先进行片段大小筛选的前提下，可用于绘制冈崎片段图谱，并检测出酿酒酵母与人类基因组中自发性单链断裂分布的偏倚与不对称性。