Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Comment
  • Published:

Advances in nanopore direct RNA sequencing

Nanopore direct RNA sequencing (DRS) reads continuous native RNA strands. Early adopters have used this technology to document nucleotide modifications and 3′ polyadenosine tails on RNA strands without added chemistry steps. Individual strands ranging in length from 70 to 26,000 nucleotides have been sequenced. In our opinion, broader acceptance of nanopore DRS by molecular biologists and cell biologists will be accelerated by higher basecall accuracy and lower RNA input requirements.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy this article

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Nanopore sequencing of individual E. coli 16S ribosomal RNA strands.
Fig. 2: General schemes for modification detection using nanopore DRS.

References

  1. Garalde, D. R. et al. Nat. Methods 15, 201–206 (2018).

    Article  CAS  Google Scholar 

  2. Viehweger, A. et al. Genome Res. 29, 1545–1554 (2019).

    Article  CAS  Google Scholar 

  3. Wongsurawat, T. et al. Front. Microbiol. 10, 260 (2019).

    Article  Google Scholar 

  4. Kim, D. et al. Cell 181, 914–921 (2020).

    Article  CAS  Google Scholar 

  5. Ugolini, C. et al. Nucleic Acids Res. 50, 3475–3489 (2022).

    Article  CAS  Google Scholar 

  6. Workman, R. E. et al. Nat. Methods 16, 1297–1305 (2019).

    Article  CAS  Google Scholar 

  7. Thomas, N. K. et al. ACS Nano 15, 16642–16653 (2021).

    Article  CAS  Google Scholar 

  8. Rousseau-Gueutin, M. et al. Gigascience 9, giaa137 (2020).

    Article  Google Scholar 

  9. Grünberger, F., Ferreira-Cerca, S. & Grohmann, D. RNA 28, 400–417 (2022).

    Article  Google Scholar 

  10. Mulroney, L. et al. RNA 28, 162–176 (2022).

    Article  CAS  Google Scholar 

  11. Sereika, M. et al. Nat. Methods 19, 823–826 (2022).

    Article  CAS  Google Scholar 

  12. Li, R. et al. Genome Res. 30, 287–298 (2020).

    Article  CAS  Google Scholar 

  13. Loman, N. J., Quick, J. & Simpson, J. T. Nat. Methods 12, 733–735 (2015).

    Article  CAS  Google Scholar 

  14. Li, H. Bioinformatics 34, 3094–3100 (2018).

    Article  CAS  Google Scholar 

  15. Tudek, A. et al. Nat. Commun. 12, 4951 (2021).

    Article  CAS  Google Scholar 

  16. Pust, M.-M., Davenport, C. F., Wiehlmann, L. & Tümmler, B. J. Bacteriol. 204, e0041821 (2022).

    Article  Google Scholar 

  17. Grünberger, F. et al. Preprint at bioRxiv https://doi.org/10.1101/2019.12.18.880849 (2020).

  18. Vo, J. M. et al. RNA 27, 1497–1511 (2021).

    Article  CAS  Google Scholar 

  19. Drexler, H. L. et al. Nat. Protoc. 16, 1343–1375 (2021).

    Article  CAS  Google Scholar 

  20. Furlan, M. et al. RNA Biol. 18, 31–40 (2021).

    Article  CAS  Google Scholar 

  21. Abebe, J. S., Verstraten, R. & Depledge, D. P. mBio 13, e0370221 (2022).

    Article  Google Scholar 

  22. White, L. K., Strugar, S. M., MacFadden, A. & Hesselberth, J. R. Preprint at bioRxiv https://doi.org/10.1101/2022.05.29.493267 (2022).

  23. Smith, A. M., Jain, M., Mulroney, L., Garalde, D. R. & Akeson, M. PLoS ONE 14, e0216709 (2019).

    Article  CAS  Google Scholar 

  24. Begik, O. et al. Nat. Biotechnol. 39, 1278–1291 (2021).

    Article  CAS  Google Scholar 

  25. Leger, A. et al. Nat. Commun. 12, 7198 (2021).

    Article  CAS  Google Scholar 

  26. Parker, M. T. et al. eLife 9, e49658 (2020).

    Article  CAS  Google Scholar 

  27. Gao, Y. et al. Genome Biol. 22, 22 (2021).

    Article  CAS  Google Scholar 

  28. Nguyen, T. A. et al. Nat. Methods 19, 833–844 (2022).

    Article  CAS  Google Scholar 

  29. Huang, S. et al. Genome Biol. 22, 330 (2021).

    Article  CAS  Google Scholar 

  30. Bailey, A. D. et al. eLife 11, e76562 (2022).

    Article  CAS  Google Scholar 

  31. Tavakoli, S. et al. Preprint at bioRxiv https://doi.org/10.1101/2021.11.03.467190 (2022).

  32. Fleming, A. M., Mathewson, N. J. & Burrows, C. J. ACS Cent. Sci. 7, 1707–1717 (2021).

    Article  CAS  Google Scholar 

  33. Pagliuca, F. W. et al. Cell 159, 428–439 (2014).

    Article  CAS  Google Scholar 

  34. Miller, R. M. et al. Genome Biol. 23, 69 (2022).

    Article  CAS  Google Scholar 

  35. Rogers, E. M. Diffusion of Innovations 5th edn (Simon and Schuster, 2003).

  36. Viscardi, M. J. & Arribere, J. A. BMC Genomics 23, 530 (2022).

    Article  CAS  Google Scholar 

  37. Pratanwanich, P. N. et al. Nat. Biotechnol. 39, 1394–1402 (2021).

    Article  CAS  Google Scholar 

  38. Parker, M. T., Barton, G. J. & Simpson, G. G. Preprint at bioRxiv https://doi.org/10.1101/2021.06.15.448494 (2021).

Download references

Acknowledgements

M.J., H.E.O. and R.A.S. were supported by NIH grant HG010053.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Miten Jain or Mark Akeson.

Ethics declarations

Competing interests

M.A. holds options in ONT and is a paid consultant to ONT. H.E.O. and M.J. received reimbursement for travel, accommodation and conference fees to speak at events organized by ONT.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jain, M., Abu-Shumays, R., Olsen, H.E. et al. Advances in nanopore direct RNA sequencing. Nat Methods 19, 1160–1164 (2022). https://doi.org/10.1038/s41592-022-01633-w

Download citation

  • Published:

  • Issue date:

  • DOI: https://doi.org/10.1038/s41592-022-01633-w

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing