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. 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N6-甲基腺苷（m6A）甲基化修饰，我们开发了一种名为GLORI的定量检测方法，可实现单碱基分辨率下的m6A修饰化学计量比检测。随后我们对施加了不同处理条件的细胞开展GLORI检测，处理方式包括细胞应激、基因敲低及抑制剂处理。后续我们进一步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修饰的功能作用。同时我们针对HEK293T、HeLa、MEF细胞及其敲低衍生细胞系的RNA测序（RNA-seq）数据开展了m6A图谱对比分析，处理条件涵盖热应激、低氧、靶向METTL3、METTL14、WTAP及YTHDF2的小干扰RNA介导的基因敲低，以及METTL3抑制剂处理。