Dissecting the effects of sequence on transcription attenuation by the Restrictor complex [TT-Seq]

The eukaryotic genome is actively transcribed by RNA Polymerase II (RNAPII) to produce both messenger RNAs (mRNAs) and a repertoire of non-coding RNAs (ncRNAs). For genome integrity, RNAPII elongation and termination is tightly regulated to enable the full-length synthesis of mRNAs and the rapid turnover of many ncRNAs. The Restrictor Complex has recently been implicated in attenuating the expression of many ncRNAs, however the mechanism by which Restrictor suppresses gene expression remains unclear. Here, we address central questions regarding the specificity of Restrictor-mediated transcription attenuation. Overall design: Examination of 11 samples by TT-seq; 3 biological replicate sets of DMSO and dTAG-13 treated cells. 2 biological replicate sets of siZC3H4 and 3 biological replicates of siNT treated cells

真核基因组可通过RNA聚合酶II（RNA Polymerase II，RNAPII）进行活跃转录，同时产生信使RNA（messenger RNAs，mRNAs）与一系列非编码RNA（non-coding RNAs，ncRNAs）。为维持基因组完整性，RNA聚合酶II的延伸与终止过程受到严格调控，以保障mRNA的全长合成，并实现多数ncRNAs的快速降解。近期研究表明，抑制因子复合物（Restrictor Complex）可参与减弱多种ncRNAs的表达，但其抑制基因表达的具体分子机制仍未明确。本研究旨在阐明该复合物介导的转录衰减特异性相关的核心科学问题。整体实验设计：采用TT-seq对11份样本进行检测；实验分组包括3批生物学重复的二甲基亚砜（DMSO）处理组细胞、3批生物学重复的dTAG-13处理组细胞、2批生物学重复的siZC3H4处理组细胞，以及3批生物学重复的非靶向小干扰RNA（siNT）处理组细胞。