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Charting the epitranscriptomic landscape across RNA biotypes using native RNA nanopore sequencing.

Gregor Diensthuber¹, Eva Maria Novoa²

¹Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, Barcelona 08003, Spain; Universitat Pompeu Fabra, Barcelona 08003, Spain.

|January 17, 2025

Summary

Direct RNA Nanopore Sequencing (DRS) revolutionizes the study of RNA modifications, enabling simultaneous identification of diverse types at single-molecule resolution. This technology offers a powerful approach to understanding the epitranscriptomic landscape across various RNA biotypes.

Keywords:

RNA modifications mRNA nanopore sequencing rRNA tRNA

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Area of Science:

Molecular Biology
Genomics
Biochemistry

Background:

RNA modifications are crucial for gene expression and cellular responses.
Studying the diverse and dynamic nature of RNA modifications presents significant challenges.
Direct RNA Nanopore Sequencing (DRS) offers a novel solution for comprehensive epitranscriptomic analysis.

Purpose of the Study:

To review the pioneering use of DRS for epitranscriptomic research.
To highlight DRS applications in investigating diverse RNA biotypes.
To discuss the future potential and challenges of DRS in RNA modification studies.

Main Methods:

Direct RNA Nanopore Sequencing (DRS) for simultaneous RNA modification identification.
Specialized library preparation protocols tailored for different RNA biotypes.
Bioinformatic workflows for detecting natural and synthetic RNA modifications.

Main Results:

DRS enables single-molecule and single-nucleotide resolution of RNA modifications.
The technology is applicable across various RNA biotypes.
DRS facilitates the detection of both endogenous and engineered RNA modifications.

Conclusions:

DRS is a transformative technology for comprehensive epitranscriptomic profiling.
Advancements in DRS chemistry are improving sequencing accuracy and yield.
Future research will leverage DRS to overcome current limitations and explore new opportunities in RNA modification biology.