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Updated: Dec 23, 2025
A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
Published on: December 5, 2016
Antibody-free enzyme-assisted chemical approach for detection of N6-methyladenosine
Ye Wang1, Yu Xiao1, Shunqing Dong1
1Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing, China.
Abstract:
The inert chemical property of RNA modification N6-methyladenosine (m6A) makes it very challenging to detect. Most m6A sequencing methods rely on m6A-antibody immunoprecipitation and cannot distinguish m6A and N6,2'-O-dimethyladenosine modification at the cap +1 position (cap m6Am). Although the two antibody-free methods (m6A-REF-seq/MAZTER-seq and DART-seq) have been developed recently, they are dependent on m6A sequence or cellular transfection. Here, we present an antibody-free, FTO-assisted chemical labeling method termed m6A-SEAL for specific m6A detection. We applied m6A-SEAL to profile m6A landscapes in humans and plants, which displayed the known m6A distribution features in transcriptome. By doing a comparison with all available m6A sequencing methods and specific m6A sites validation by SELECT, we demonstrated that m6A-SEAL has good sensitivity, specificity and reliability for transcriptome-wide detection of m6A. Given its tagging ability and FTO's oxidation property, m6A-SEAL enables many applications such as enrichment, imaging and sequencing to drive future functional studies of m6A and other modifications.
Insights
A new method, m6A-SEAL, enables specific detection of RNA N6-methyladenosine (m6A) modifications without antibodies. This antibody-free approach offers reliable transcriptome-wide m6A profiling for future functional studies.
Area of Science:
- Molecular Biology
- Epigenetics
- RNA Biology
Background:
- N6-methyladenosine (m6A) is a crucial RNA modification, but its detection is challenging due to its inert chemical properties.
- Existing m6A sequencing methods often rely on antibodies and struggle to differentiate between m6A and cap m6Am, or have limitations like sequence dependence or cellular transfection requirements.
Purpose of the Study:
- To develop a novel, antibody-free method for specific and reliable detection of m6A modifications.
- To profile m6A landscapes in humans and plants using the new method.
- To demonstrate the sensitivity, specificity, and reliability of the new method compared to existing techniques.
Main Methods:
- Development of m6A-SEAL, an antibody-free chemical labeling method assisted by FTO enzyme.
- Application of m6A-SEAL for transcriptome-wide m6A profiling in human and plant samples.
- Comparative analysis of m6A-SEAL with existing m6A sequencing methods and validation using SELECT.
Main Results:
- m6A-SEAL successfully profiled m6A landscapes in humans and plants, aligning with known distribution patterns.
- The method demonstrated good sensitivity, specificity, and reliability in transcriptome-wide m6A detection.
- Validation confirmed the accuracy of m6A-SEAL in identifying specific m6A sites.
Conclusions:
- m6A-SEAL provides a robust, antibody-free solution for specific m6A detection.
- The method's ability to tag m6A modifications opens avenues for enrichment, imaging, and sequencing.
- m6A-SEAL is poised to advance functional studies of m6A and other RNA modifications.
