Improvements in nanopore RNA sequencing and how they affect biological research

Nanopore direct RNA sequencing (DRS) enables full-length, single-molecule sequencing of native RNA, capturing transcript isoforms and preserving modifications without cDNA conversion.

• DRS facilitates the analysis of various RNA types, including mRNA, rRNA, tRNA, circRNA, and viral RNA.
• It allows for detailed quantification of transcript isoforms, measurement of poly(A) tails, and identification of fusion transcripts.
• The technology has been instrumental in discovering novel transcripts and alternative splicing events in cancer, plants, and parasites.
• DRS enables the direct detection of RNA modifications such as m6A, m5C, pseudouridine, and RNA editing events.
• Emerging applications include co-transcriptional splicing analysis, lncRNA and circRNA discovery, and real-time RNA structural mapping.
• Despite limitations in sequencing accuracy and cost, ongoing improvements in technology may enhance DRS's utility in transcriptome profiling and RNA regulatory analysis.

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