Genome-wide analysis of mRNAs expression in prometaphase

The goal of this experiment was to compare the transcriptome of cells that overexpressed the RNA-binding protein Staufen1 to that of cells that expressed the empty vector as control. Stau1 abundance fluctuates through the cell cycle: it is high in the G2 phase of the cell cycle and rapidly decreases as cells transit through mitosis. The importance of controlling Stau1 levels was underscored by the observation that its overexpression impairs mitosis as well as proliferation of transformed cell lines. As a major post-transcriptional regulator, Stau1 may exert its role(s) through the spatial and/or temporal regulation of its bound mRNAs. Therefore, to gather clues to support this possibility, we determined if Stau1 overexpression significantly modified the transcriptome of transformed cells in prometaphase explaining the observed impairment in mitosis transit and/or cell proliferation. HEK293T cells were transfected with plasmids coding for Stau155-FLAG or the empty vector as control. Cells were synchronized in prometaphase with nocodazole.