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Updated: May 28, 2026
08:56
A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
Epitranscriptomic Stability-Variable Extents of N1-Methyladenosine to N6-Methyladenosine Conversion Under Different
Frank Morales Shnaider1, Hasna Kanan1, Shrikant Patel1
1Department of Chemistry and Biochemistry, University of North Carolina Greensboro, 301 McIver St., Greensboro, NC 27412, USA.
Biomolecules
|May 27, 2026
Summary
RNA modification stability is crucial for epitranscriptome research. N1-methyladenosine (m1A) converts to N6-methyladenosine (m6A) under various conditions, impacting quantification accuracy and requiring careful experimental design.
Area of Science:
- Molecular Biology
- Biochemistry
- Epigenetics
Background:
- The epitranscriptome, encompassing RNA modifications, is a rapidly expanding field.
- Accurate quantification of RNA modifications is essential for understanding their biological roles.
- The stability of RNA modifications under standard experimental conditions is not well understood.
Purpose of the Study:
- To investigate the stability of N1-methyladenosine (m1A) under common RNA experimental conditions.
- To identify factors influencing m1A stability and potential conversion products.
- To propose a method for mitigating quantification bias caused by m1A instability.
Main Methods:
- Exposure of m1A ribonucleoside to various pH, temperature, and concentration conditions.
- Analysis of samples using untargeted high-resolution mass spectrometry.
- Assessment of m1A conversion in a biological sample and application of the standard addition method.
Main Results:
- m1A converts to N6-methyladenosine (m6A) under alkaline, neutral, and elevated temperature conditions.
- The rate of m1A to m6A conversion is dependent on initial m1A concentration.
- Non-enzymatic deamination of m1A to N1-methylinosine was detected in trace amounts.
- m1A to m6A conversion was observed in biological samples, necessitating the standard addition method for accurate quantification.
Conclusions:
- The stability of m1A is compromised under specific experimental conditions, leading to m1A-to-m6A conversion.
- Variations in m1A concentration can significantly affect conversion rates, posing challenges for comparative studies.
- The standard addition method provides a viable solution to correct for m1A-to-m6A conversion bias in quantification.
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