Absolute quantification of single-base m6A methylation in the mammalian transcriptome

To investigate the m6A methylation in the mammalian transcriptome, we developed a quantitative method, names GLORI, that could detect m6A stoichiometry at single-base resolution. We then performed GLORI on cells with different treatment, such as stress, knockdown and inhibitor. We next analysis m6A methyloms of different celllines and cells under different treatment to investigate the fuctional role of m6A. Overall design: Comparation m6A profiling analysis of RNA-seq data for HEK293T/HeLa/MEF/cells and its KD derivatives (Heatshock, hypoxia, siMETTL3, siMETTL14, siWTAP, siYTHDF2, and inhibition of METTL3).

为探究哺乳动物转录组中的m6A甲基化修饰，我们开发了一种名为GLORI的定量检测方法，可在单碱基分辨率下实现m6A化学计量比的检测。随后，我们对施加了不同处理的细胞开展GLORI实验，处理方式涵盖应激处理、基因敲低与抑制剂处理。接下来，我们分析了不同细胞系以及经不同处理的细胞的m6A甲基化组，以探究m6A的生物学功能。整体实验设计：对HEK293T、HeLa、MEF细胞及其基因敲低（KD）衍生细胞（热休克、缺氧、siRNA靶向METTL3、siRNA靶向METTL14、siRNA靶向WTAP、siRNA靶向YTHDF2，以及METTL3抑制剂处理组）的RNA测序（RNA-seq）数据开展m6A谱图比较分析。