Institute of Molecular Biology (IMB) gGmbH, Mainz, Germany.
FEBS J. 2021 Jul;288(13):3948-3961. doi: 10.1111/febs.15568. Epub 2020 Oct 2.
Mapping the genome-wide distribution of DNA lesions is key to understanding damage signalling and DNA repair in the context of genome and chromatin structure. Analytical tools based on high-throughput next-generation sequencing have revolutionized our progress with such investigations, and numerous methods are now available for various base lesions and modifications as well as for DNA double-strand breaks. Considering that single-strand breaks are by far the most common type of lesion and arise not only from exposure to exogenous DNA-damaging agents, but also as obligatory intermediates of DNA replication, recombination and repair, it is surprising that our insight into their genome-wide patterns, that is the 'SSBreakome', has remained rather obscure until recently, due to a lack of suitable mapping technology. Here we briefly review classical methods for analysing single-strand breaks and discuss and compare in detail a series of recently developed high-resolution approaches for the genome-wide mapping of these lesions, their advantages and limitations and how they have already provided valuable insight into the impact of this type of damage on the genome.
绘制全基因组范围内 DNA 损伤的图谱对于理解基因组和染色质结构背景下的损伤信号转导和 DNA 修复至关重要。基于高通量下一代测序的分析工具极大地推动了这些研究的进展，现在有许多方法可用于各种碱基损伤和修饰以及 DNA 双链断裂。鉴于单链断裂迄今为止是最常见的损伤类型，不仅来自于外源性 DNA 损伤剂的暴露，而且还是 DNA 复制、重组和修复的必需中间体，令人惊讶的是，由于缺乏合适的图谱技术，我们对其全基因组图谱（即“SSBreakome”）的了解直到最近才变得相当模糊。在这里，我们简要回顾了分析单链断裂的经典方法，并详细讨论和比较了一系列最近开发的用于全基因组图谱绘制这些损伤的高分辨率方法，以及它们的优点和局限性，以及它们如何已经为这种类型的损伤对基因组的影响提供了有价值的见解。
