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Updated: Jun 24, 2026
08:45
Characterizing RNA Modifications in Single Neurons Using Mass Spectrometry
Published on: April 21, 2022
Quantitative profiling of RNA modifications enriched in non-membrane-bound cellular structures using APEX-RNA-MS
Kyung W Seo1, Dhruv Y Dhingani1, Ralph E Kleiner1
1Department of Chemistry, Princeton University, Princeton, NJ 08544, USA.
Cell Chemical Biology
|June 22, 2026
Summary
This study introduces APEX-RNA-MS, a new method to map RNA modifications within cells. It reveals how RNA modifications change in cellular structures during stress, offering insights into RNA biology.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- RNA modifications are crucial for biological processes.
- Understanding their subcellular distribution is challenging.
- Post-transcriptional regulation controls RNA function.
Purpose of the Study:
- To develop a method for characterizing RNA modifications at subcellular resolution.
- To investigate the distribution of RNA modifications near RNA-binding proteins in cellular condensates.
- To analyze changes in RNA modifications during cellular stress responses.
Main Methods:
- APEX-RNA-MS combines APEX2 proximity labeling with LC-MS.
- Quantification of modified ribonucleotides.
- RNA sequencing and RNA FISH were used for validation.
Main Results:
- APEX-RNA-MS successfully mapped RNA modifications proximal to proteins in non-membrane-bound structures.
- Changes in protein-proximal RNA modification levels were observed upon DNA damage and stress granule formation.
- tRNA-specific modifications were found proximal to G3BP1, with tRNAs accumulating in stress granules.
Conclusions:
- APEX-RNA-MS provides a versatile approach for studying subcellular RNA modification distribution.
- The study offers new insights into the composition and function of cellular condensates.
- Characterizing RNA modifications in dynamic cellular structures is now more feasible.
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