Genome-wide distribution of transcription-coupled nucleotide excision repair and RNA polymerase II in DRB-treated XP-C cells after UV damage

The fate of RNAPII in the course of the transcription-coupled repair pathway is unclear. To address this problem we have used methods to control transcription, so as to initiate a discrete 'wave' of elongation complexes. We also used methods to identify where elongation complexes and transcription-repair coupling events are located in genes throughout the genome. Overall design: XP-C cells which is deficient in the global repair were treated with different DRB regimens. After UV irradiation, XR-seq and mNET were performed to analyze the repair and the localization of elongating RNA polymerase II

RNA聚合酶II（RNAPII）在转录偶联修复（transcription-coupled repair, TCR）通路中的动态去向尚未明确。为解决这一科学问题，本研究采用转录调控方法，诱导形成离散的延伸复合物‘波’。同时，我们建立了特异性检测手段，以定位全基因组范围内基因内部的延伸复合物与转录-修复偶联事件的分布位点。
实验整体设计：选取全局修复缺陷的XP-C细胞，通过不同DRB处理方案进行干预。经紫外线（UV）辐照后，借助XR-seq与mNET技术分别检测修复进程，并分析延伸态RNA聚合酶II的定位特征。