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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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Quantitative analysis of m6A RNA modification by LC-MS
Lavina Mathur1, Sunhee Jung2, Cholsoon Jang2
1Department of Microbiology and Molecular Genetics, Chao Family Comprehensive Cancer Center, University of California Irvine School of Medicine, Irvine, CA 92697-4025, USA.
STAR Protocols
|August 17, 2021
Summary
This study presents a new protocol to measure N6-adenosine methylation (m6A) levels in messenger RNA (mRNA). Accurate m6A quantification is vital for understanding gene expression regulation.
Area of Science:
- Molecular Biology
- Biochemistry
- Epigenetics
Background:
- N6-adenosine methylation (m6A) is a crucial post-transcriptional modification of messenger RNA (mRNA).
- m6A regulates fundamental gene expression processes.
- Accurate quantification of m6A levels is essential for biological research.
Purpose of the Study:
- To provide a detailed protocol for quantifying adenosine and m6A levels in cellular mRNAs.
- To enable researchers to accurately measure m6A dynamics in various biological contexts.
Main Methods:
- The protocol involves digesting mRNA into nucleosides using nucleases and phosphatases.
- Quantification of nucleosides is performed using liquid chromatography-mass spectrometry (LC-MS).
Main Results:
- The developed protocol allows for the precise measurement of both adenosine and m6A levels in mRNA.
- This method facilitates the accurate assessment of m6A stoichiometry in cellular mRNA populations.
Conclusions:
- This protocol offers a reliable method for quantifying m6A levels in mRNA.
- Accurate m6A measurement is critical for advancing the understanding of gene regulation and its role in biological systems.
