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Updated: Nov 15, 2025
05:41
2D-HELS MS Seq: A General LC-MS-Based Method for Direct and de novo Sequencing of RNA Mixtures with Different Nucleotide Modifications
Published on: July 10, 2020
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Analysis of RNA Modifications by Second- and Third-Generation Deep Sequencing: 2020 Update
Yuri Motorin1, Virginie Marchand2
1Université de Lorraine, CNRS, IMoPA (UMR7365), F54000 Nancy, France.
Genes
|March 6, 2021
Summary
Mapping RNA modifications is crucial in epitranscriptomics. New sequencing and nanopore technologies offer advanced methods for detecting and quantifying these vital RNA changes.
Area of Science:
- Biochemistry
- Molecular Biology
- Genomics
Background:
- Accurate mapping and quantification of RNA modifications are essential in epitranscriptomics.
- Numerous RNA modifications exist across various RNA types, including mRNA, tRNA, rRNA, and ncRNA/miRNA.
- The discovery of N6-methyladenosine (m6A) in mRNA spurred the development of detection methods.
Purpose of the Study:
- To review recent advancements in mapping and quantifying RNA modifications.
- To highlight novel deep sequencing and direct RNA sequencing approaches.
- To discuss the challenges and progress in the field of epitranscriptomics.
Main Methods:
- Deep sequencing-based methods utilizing reverse transcription signatures.
- Chemical or enzymatic treatments to induce detectable signatures.
- Labeling RNA at abasic sites followed by adapter ligation.
- Affinity/immunoprecipitation protocols using antibodies or specific binding proteins.
- Direct single-molecule RNA sequencing using nanopores.
Main Results:
- Significant expansion of detectable modified RNA residues.
- Development of diverse sequencing and labeling strategies.
- Emerging potential of nanopore sequencing for direct RNA modification mapping.
Conclusions:
- The field of RNA modification analysis is rapidly evolving.
- Advanced sequencing technologies are crucial for comprehensive epitranscriptomic studies.
- Direct RNA sequencing holds promise for real-time, high-resolution mapping of RNA modifications.
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