IFOM (Fondazione Istituto FIRC di Oncologia Molecolare), Milan, Italy.
Centre for DNA Fingerprinting and Diagnostics, Hyderabad, Telangana, India.
Methods Mol Biol. 2022;2528:317-328. doi: 10.1007/978-1-0716-2477-7_21.
DNA hybrids are generated naturally behind the elongating RNA polymerase as a transcriptional intermediate. However, prolonged persistence of these structures challenges the integrity of the genome by creating R-loops and by interfering with DNA replication and other chromatin related processes. Precise mapping and characterization of their distribution across the genome has been a major challenge to understand the genesis of RNA:DNA hybrids and their conversion into genotoxic intermediates. Here we provide the detailed protocol for mapping RNA:DNA hybrid across the Saccharomyces cerevisiae genome.
DNA 杂交体作为转录中间体，在延伸的 RNA 聚合酶后面自然产生。然而，这些结构的长时间持续存在会通过形成 R 环并干扰 DNA 复制和其他与染色质相关的过程，对基因组的完整性构成挑战。精确绘制和描述它们在基因组中的分布情况，一直是理解 RNA:DNA 杂交体的起源及其转化为遗传毒性中间产物的主要挑战。在这里，我们提供了在酿酒酵母基因组中绘制 RNA:DNA 杂交体的详细方案。
