Functional studies and biogenesis of RNA modifications

本项目旨在研究与各种生物过程相关的RNA分子转录后修饰的功能和生物发生。我们使用RNA质谱仪和反向遗传学相结合的方法，对负责RNA修饰的生物发生的基因进行了系统的全基因组筛选。我们发现了负责合成2-硫代尿苷、乳酪氨酸、赖氨酸和N4-乙酰胞苷的新基因，这些基因存在于大肠杆菌或酿酒酵母的tRNA中。为了识别人脑mRNAs中的肌苷，我们建立了一种名为肌苷化学擦除(ICE)的生化方法，直接检测RNA链上的肌苷。我们已经绘制了超过16,000个肌苷在人脑转录组中的新位置。此外，我们还启动了一项利用ICE方法结合深度测序(ICE-Seq)对肌苷位点进行全基因组鉴定的项目。

This project aimed to study on functional aspects and biogenesis of post-transcriptional modifications of RNA molecules associated with various biological processes. We employed a systematic genome-wide screen of genes responsible for biogenesis of RNA modifications using RNA mass spectrometry combined with reverse genetic approach. We identified novel genes responsible for synthesizing 2-thiouridine, wybutosine, lysidine and N4-acetylcytidine that are found in tRNAs from E. coli or S. cerevisiae. For identification of inosines in mRNAs from human brain, we have established a biochemical method, named "inosine chemical erasing (ICE)", to directly detect inosines on RNA strands. We have mapped more than 16,000 novel sites of inosines in human brain transcriptome. Furthermore, we have started a project of genome-wide identification of inosine sites by using the ICE-method combined with the deep sequencing (ICE-Seq).