1. Academic Validation
  2. Absolute quantification of single-base m6A methylation in the mammalian transcriptome using GLORI

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

  • Nat Biotechnol. 2022 Oct 27. doi: 10.1038/s41587-022-01487-9.
Cong Liu # 1 Hanxiao Sun # 1 Yunpeng Yi # 2 3 Weiguo Shen # 2 Kai Li # 4 5 Ye Xiao 4 5 Fei Li 2 Yuchen Li 4 5 Yongkang Hou 2 Bo Lu 1 Wenqing Liu 6 7 Haowei Meng 1 Jinying Peng 1 Chengqi Yi 8 9 10 Jing Wang 11
Affiliations

Affiliations

  • 1 State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China.
  • 2 State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, China.
  • 3 Shandong Provincial Animal and Poultry Green Health Products Creation Engineering Laboratory, Institute of Poultry Science, Shandong Academy of Agricultural Science, Jinan, China.
  • 4 Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China.
  • 5 Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China.
  • 6 School of Life Sciences, Tsinghua University, Beijing, China.
  • 7 Tsinghua-Peking Joint Center for Life Sciences, Tsinghua University, Beijing, China.
  • 8 State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China. chengqi.yi@pku.edu.cn.
  • 9 Peking-Tsinghua Center for Life Sciences, Peking University, Beijing, China. chengqi.yi@pku.edu.cn.
  • 10 Department of Chemical Biology and Synthetic and Functional Biomolecules Center, College of Chemistry and Molecular Engineering, Peking University, Beijing, China. chengqi.yi@pku.edu.cn.
  • 11 State Key Laboratory of Natural and Biomimetic Drugs, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Beijing, China. wangjingsioc@pku.edu.cn.
  • # Contributed equally.
Abstract

N6-methyladenosine (m6A) is the most abundant RNA modification in mammalian cells and the best-studied epitranscriptomic mark. Despite the development of various tools to map m6A, a transcriptome-wide method that enables absolute quantification of m6A at single-base resolution is lacking. Here we use glyoxal and nitrite-mediated deamination of unmethylated adenosines (GLORI) to develop an absolute m6A quantification method that is conceptually similar to bisulfite-sequencing-based quantification of DNA 5-methylcytosine. We apply GLORI to quantify the m6A methylomes of mouse and human cells and reveal clustered m6A modifications with differential distribution and stoichiometry. In addition, we characterize m6A dynamics under stress and examine the quantitative landscape of m6A modification in gene expression regulation. GLORI is an unbiased, convenient method for the absolute quantification of the m6A methylome.

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