The functional impact of RNA Polymerase II pausing across multiple mamamlian cell types. Homo sapiens

We analyzed the breadth and functional relationships of RNA Polymerase II pausing across many human and mouse cell types to understand what roles RNA Polymerase II pausing plays in gene regulation. We identified a novel association of H2A.Z at the TSS of increasingly paused genes. We knocked down H2A.Z to test whether H2A.Z positively or negatively affects RNA Polymerase II pausing to find that pausing globally increased upon knockdown. Overall design: We tested the effect of H2A.Z knockdown on RNA Polymerase II pausing by setting up a case vs. control experiment in which MCF7 breast cancer cells were treated with a siRNA against H2A.Z. We tested for changes in RNA Polymerase II pausing as a result.

本研究针对多种人类与小鼠细胞类型中的RNA聚合酶II（RNA Polymerase II）暂停现象的广度及功能关联展开系统分析，旨在阐明RNA聚合酶II暂停在基因调控过程中所发挥的作用。我们在转录暂停程度逐渐升高的基因的转录起始位点（Transcription Start Site, TSS）处，鉴定出组蛋白H2A.Z（H2A.Z）的全新关联特征。为验证H2A.Z对RNA聚合酶II暂停存在正向还是负向调控作用，我们通过敲低H2A.Z的表达开展功能验证实验，结果发现敲低后全局范围内的RNA聚合酶II暂停现象均显著增强。整体实验设计：本研究以MCF7乳腺癌细胞为实验模型，设置实验组与对照组，将MCF7细胞分别用靶向H2A.Z的小干扰RNA（small interfering RNA, siRNA）及阴性对照siRNA处理，以此检测H2A.Z敲低所引发的RNA聚合酶II暂停相关变化。