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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
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Absolute quantification of single-base m6A methylation in the mammalian transcriptome using GLORI
Cong Liu1, Hanxiao Sun1, Yunpeng Yi2,3
1State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing, China.
Nature Biotechnology
|October 27, 2022
Summary
A new method called Glori allows precise, whole-transcriptome measurement of N6-methyladenosine (m6A) RNA modifications. This breakthrough enables absolute quantification of m6A, advancing epitranscriptomic research in mammals.
Area of Science:
- Epitranscriptomics
- Molecular Biology
- Genomics
Background:
- N6-methyladenosine (m6A) is the most prevalent RNA modification in mammals.
- Existing methods for m6A mapping lack transcriptome-wide absolute quantification at single-base resolution.
Purpose of the Study:
- To develop a novel method for absolute quantification of m6A modifications across the transcriptome.
- To apply this method to analyze m6A methylomes in mammalian cells.
Main Methods:
- Development of the Glyoxal and Nitrite-mediated Deamination of Unmethylated Adenosines (GLORI) technique.
- GLORI is analogous to bisulfite sequencing for DNA methylation quantification.
- Application of GLORI to mouse and human cell methylomes.
Main Results:
- GLORI enables absolute quantification of m6A at single-base resolution.
- Revealed clustered m6A modifications with distinct distribution and stoichiometry in mouse and human cells.
- Characterized m6A dynamics under stress and its role in gene expression regulation.
Conclusions:
- GLORI provides an unbiased and convenient method for absolute m6A methylome quantification.
- The study offers new insights into the quantitative landscape and dynamics of m6A modifications.
