The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [RNA-seq]

We performed high-throughput mRNA sequencing during a meiotic time course (0, 2, 3, 5, 6, 7 and 9 hr) in WT, pho92 mutant and ime4 mutant cells. Overall design: Meiosis was induced by resuspension of cells in SPO medium and high synchronicity was achieved thanks to the SK1 pCUP-IME1 background alowing inducible activation of IME1 after 2 hr in SPO by addition of 50 µM of copper (II) sulfate. PolyA RNAs were extracted from cells collected at 7 time points from 3 different strains (WT, pho92 mutant, ime4 mutant) and in 3 biological replicates, giving a total of 63 yeast mRNA-seq samples.

本研究针对野生型（Wild Type, WT）、pho92突变体与ime4突变体酵母细胞，在减数分裂时间进程（0、2、3、5、6、7和9小时）中开展了高通量mRNA测序。整体实验设计如下：通过将细胞重悬于SPO培养基中诱导减数分裂；依托SK1 pCUP-IME1遗传背景，可在SPO培养基培养2小时后，通过添加50 μM硫酸铜（II）诱导激活IME1基因，以此实现高同步化的减数分裂进程。本研究从3株不同菌株的7个时间点收集的细胞中提取聚腺苷酸RNA，并设置3次生物学重复，最终共计获得63个酵母mRNA测序样本。