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Absolute quantification of single-base m6A methylation in the mammalian transcriptome using GLORI
Abstract
N 6 -methyladenosine (m 6 A) is the most abundant RNA modification in mammalian cells and the best-studied epitranscriptomic mark. Despite the development of various tools to map m 6 A, a transcriptome-wide method that enables absolute quantification of m 6 A at single-base resolution is lacking. Here we use glyoxal and nitrite-mediated deamination of unmethylated adenosines (GLORI) to develop an absolute m 6 A quantification method that is conceptually similar to bisulfite-sequencing-based quantification of DNA 5-methylcytosine. We apply GLORI to quantify the m 6 A methylomes of mouse and human cells and reveal clustered m 6 A modifications with differential distribution and stoichiometry. In addition, we characterize m 6 A dynamics under stress and examine the quantitative landscape of m 6 A modification in gene expression regulation. GLORI is an unbiased, convenient method for the absolute quantification of the m 6 A methylome.
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“GLORI was performed as reported previously (Liu et al., 2023), slightly modified, as follows:”
From METTL3 uncouples chromatin accessibility from transcription during retinal development (2025) · cites this paper as Liu et al., 2023
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Metadata
- Authors
- Liu, C., Sun, H., Yi, Y., Shen, W., Li, K., Xiao, Y., Li, F., Li, Y., Hou, Y., Lu, B., Liu, W., Meng, H., Peng, J., Yi, C., & Wang, J.
- Affiliations
- Peking University (CN)
- Center for Life Sciences (CN)
- Corresponding author
- Yi, Chengqi
- Wang, Jing
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2022-10-27
- Volume
- 41
- Issue
- 3
- Pages
- 355-366
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W4307500724
- Cited by
- 386
- References
- 67
- Citations by year
- 2026: 100 · 2025: 125 · 2024: 103 · 2023: 53 · 2022: 4
- FWCI
- 23.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Natural Science Foundation of China (22077006, 91940304)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Keywords
- N6-methyladenosine, m6A RNA methylation, epitranscriptomics, absolute quantification, single-base resolution, m6A methylome, stress response, gene expression regulation, mouse cells, human cells
- MeSH terms
- Animals, DNA, Gene Expression Regulation, Humans, Mammals, Methylation, DNA Methylation, Transcriptome
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