Genome-wide Map of R-loop Reveals its interplays with Transcription and Genome Integrity during Germ Cell Meiosis

R-loop is a three-strand nucleic acid structure composed of mRNA and DNA. In mammalian meiotic cells, R-loop mainly distributed at promoter-related regions, and combined with systematically transcriptome analysis showed R-loop was closely related to transcription in meiotic process. Moreover, knockout of Rnaseh1 in meiotic cells caused mice completely sterile with R-loop accumulation and abnormal DSB repairing in spermatocytes. Further analysis showed R-loop accumulation in leptotene/zygotene influenced transcriptional regulation in pachytene stage. Thus, mutual regulation of R-loop and transcription plays essential role in spermatogenesis. A total of 12 samples were analyzed in this study, including WT and Rnaseh1 knock-out spermatogenic cells from two stages, leptotene/zygotene and pachytene. Each biological samples has three replicates.R环（R-loop）是由信使RNA（mRNA）与DNA构成的三链核酸结构。在哺乳动物减数分裂细胞中，R环主要分布于启动子相关区域；结合系统性转录组分析结果显示，R环与减数分裂过程中的转录活动密切相关。此外，在减数分裂细胞中敲除核糖核酸酶H1（Rnaseh1）会导致小鼠完全不育，同时精母细胞内出现R环积累与DNA双链断裂（DSB）修复异常。进一步分析表明，细线期/偶线期的R环积累会影响粗线期的转录调控。因此，R环与转录的相互调控在精子发生过程中发挥关键作用。本研究共分析12份样本，涵盖细线期/偶线期、粗线期两个发育阶段的野生型（Wild Type, WT）与Rnaseh1敲除生精细胞，每份生物学样本设置3次生物学重复。