3'UTR-dependent activity of transcription factors and chromatin regulators [RNA-seq]

Over 2700 human mRNAs have highly conserved 3'UTRs, but their biological functions are unknown. These mRNAs enrich for transcriptional regulators. Therefore, we performed RNA-seq to determine the roles of 3'UTRs in controlling the cellular activity of selected transcriptional regulators, here MYC and UTX. Overall design: RNA-seq profiling of U2OS or MCF7 cell lines transfected with cDNA vectors encoding the transcription factor MYC or the histone demethylase UTX. Gene expression profiles were compared to control, after each protein was expressed from an mRNA template that only contained the coding sequence (-NU) or that contained the coding sequence and its full-length 3'UTR (-U).

已有超过2700种人类信使RNA（mRNA）拥有高度保守的3'非翻译区（3'UTR），但其生物学功能尚未明确。该类信使RNA显著富集于转录调控因子类群。因此，本研究通过RNA测序（RNA-seq）探究3'UTR在调控所选转录调控因子（此处为MYC与UTX）的细胞活性中的作用。实验整体设计：对转染了编码转录因子MYC或组蛋白去甲基化酶UTX的互补DNA（cDNA）载体的U2OS与MCF7细胞系进行RNA-seq表达谱分析。将各组蛋白分别由仅含编码序列的mRNA模板（-NU组）或携带有编码序列及其全长3'UTR的mRNA模板（-U组）表达后，对比其基因表达谱与对照组的差异。