DEtail-seq profiled meiotic DSBs in Homo sapiens. DEtail-seq profiled meiotic DSBs in Homo sapiens

SPO11-promoted DNA double-strand breaks (DSBs) formation is a crucial step for meiotic recombination, and it is indispensable to detect the broken DNA ends accurately for dissecting the molecular mechanisms behind. Here, we report a novel technique, named DEtail-seq (DNA End tailing followed by sequencing), that can directly and quantitatively capture the meiotic DSB 3’ overhang hotspots at single-nucleotide resolution. Overall design: Adult human testis samples were obtained from two patients with obstructive azoospermia (OA) who underwent vasoepididymostomy. The patients with OA were enrolled from Reproductive Center of Peking University Third Hospital. We excluded patients with chromosomal abnormalities, Y-chromosome microdeletions, varicocele, metabolic disorders such as diabetes and hepatic disease, occupational exposure to the agents, and other known factors related to male infertility.

SPO11介导的DNA双链断裂（DNA double-strand breaks, DSBs）形成是减数分裂重组的关键步骤，精准检测断裂DNA末端对于解析其背后的分子机制不可或缺。本研究报道一种名为DEtail-seq（DNA末端加尾后测序，DNA End tailing followed by sequencing）的新型技术，该技术能够以单核苷酸分辨率直接、定量地捕获减数分裂DSB的3'突出端热点区域。整体实验设计：成年人类睾丸样本取自2名接受输精管附睾吻合术的梗阻性无精子症（obstructive azoospermia, OA）患者，上述OA患者招募自北京大学第三医院生殖医学中心。本研究排除了存在染色体异常、Y染色体微缺失、精索静脉曲张、糖尿病及肝脏疾病等代谢紊乱、职业性接触有害因子，以及其他已知男性不育相关因素的患者。