Evidence map›Paper›PMID 41325774›Full record

ArticleNucleic acids research2025

Direct RNA sequencing enables improved transcriptome assessment and tracking of RNA modifications for medical applications.

Charlotte Hewel, Anna Wierczeiko, Johannes Miedema, Johannes Friedrich, Felix Hofmann, Stephan Weißbach, Vincent Dietrich, Laura Holthöfer, Verena Haug, Stefan Mündnich and 16 more

Abstract read
In one paragraph

Article in Nucleic acids research, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.

0numbers the graph read from it
0cells of the map it votes in
22citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

22 citing papers in PubMed.

  1. Review
  2. Article
  3. Compendium of RNA modifications for bacterial stress adaptation.Microbiology and molecular biology reviews : MMBR · 2026
    Review
  4. Article
  5. Primer-Less Species Identification Throughout Fungal (International journal of molecular sciences · 2026
    Article
  6. Epitranscriptomic Analysis of A-to-I RNA Editing and mInternational journal of molecular sciences · 2026
    Review
  7. Article
  8. Integrating mass spectrometry with Nanopore direct RNA sequencing forbioRxiv : the preprint server for biology · 2026
    Article
  9. Article
  10. Review
  11. Article
  12. Article
  13. The bladder cancer mEMBO reports · 2026
    Article
  14. Article
  15. Article
  16. Review
  17. Review
  18. Article
  19. FTO depletion does not alter mbioRxiv : the preprint server for biology · 2025
    Article
  20. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

26 authors.

Charlotte HewelInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Anna WierczeikoInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Johannes MiedemaInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Johannes FriedrichInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Felix HofmannInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Stephan WeißbachInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.ORCID 0009-0005-4347-952X
Vincent DietrichInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Laura HolthöferInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Verena HaugDepartment for Pediatrics, University Medical Center of the Johannes Gutenberg University Mainz, 55131 Mainz, Germany.
Stefan MündnichInstitute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University Mainz, Mainz 55128, Germany.
Lukas SchartelBiocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, Mainz 55128, Germany.
Lioba LehmannInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Kristi L JensenBiocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, Mainz 55128, Germany.
Stefan DiederichInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Stanislav SysInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Tamer ButtoInstitute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University Mainz, Mainz 55128, Germany.ORCID 0000-0001-8028-0038
Norbert W PaulInstitute for the History, Philosophy, and Ethics of Medicine, Johannes Gutenberg University Medical Center Mainz, Mainz 55131, Germany.
Jonas KochDivision of Epigenetics, DKFZ-ZMBH Alliance, German Cancer Research Center, Heidelberg 69120, Germany.
Frank LykoDivision of Epigenetics, DKFZ-ZMBH Alliance, German Cancer Research Center, Heidelberg 69120, Germany.
Florian KraftInstitut für Humangenetik und Genommedizin, Uniklinik RWTH Aachen, Aachen, Germany.ORCID 0000-0002-5324-9155
Alexandra RussoDepartment for Pediatrics, University Medical Center of the Johannes Gutenberg University Mainz, 55131 Mainz, Germany.ORCID 0009-0000-1141-7977
Susann SchweigerInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Edward A LemkeBiocenter, Johannes Gutenberg University Mainz, Hanns-Dieter-Hüsch-Weg 17, Mainz 55128, Germany.ORCID 0000-0002-0634-0503
Mark HelmInstitute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University Mainz, Mainz 55128, Germany.ORCID 0000-0002-0154-0928
Matthias LinkeInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.
Susanne GerberInstitute for Human Genetics, University Medical Center of the Johannes Gutenberg University Mainz, Mainz 55131, Germany.ORCID 0000-0001-9513-0729

Funding

Boehringer Ingelheim StiftungCarl-Zeiss-StiftungDeutsche ForschungsgemeinschaftDFG 464588647ERCGerman Research Foundation 439669440 TRR319 RMaP TP A01/A05/C01/C03/C04IQCB
6 · The paper itself

Abstract

Direct RNA sequencing (DRS) is a Nanopore-based technique for analyzing RNA in its native form. This technique promises breakthroughs in diagnostics and biomarker development. Coupled to RNA002 sequencing chemistry, its clinical implementation has been challenging due to low throughput, low accuracy, and lack of large-scale RNA-modification models. In this study, we evaluate the improvements achieved by pairing the latest RNA004 chemistry with novel modified-base-calling models for pseudouridine and N6-methyladenosine using diverse RNA samples from cell lines, synthetic oligos, and human blood. Finally, we present the first clinical application of DRS by confirming the loss of RNA methylation in a patient carrying truncating mutations in the methyltransferase METTL5. Conclusively, the combined use of RNA004 chemistry with the base-calling models significantly improved the throughput, accuracy, and site-specific detection of modifications. From this perspective, we offer an outlook on the potential suitability of DRS for use in routine diagnostics, as well as the first comprehensive benchmark of human peripheral blood. Furthermore, we demonstrate on the basis of a stop-codon readthrough enhancing agent potential roadblocks for routine quality assessments of RNA therapeutics.

Indexed as

RNARNA Processing, Post-TranscriptionalSequence Analysis, RNATranscriptomeAdenosineHigh-Throughput Nucleotide SequencingHumansMethylationMethyltransferasesPseudouridineAdenosineMethyltransferasesN-methyladenosinePseudouridineRNA

Identifiers

PMID41325774
PMCPMC12663090

What Socratic holds

Textmetadata
LicenceCC BY-NC
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the Socratic graph.