2024/07/25 by Charlotte Hewel, Anna Wierczeiko, Johannes Miedema +22 · 1 voice · 1 citation
Biochemistry, Genetics and Molecular Biology · #RNA modifications and cancer #Cancer-related molecular mechanisms research #MicroRNA in disease regulation
paper · pdf · doi:10.1101/2024.07.25.605188
openalex publication_date 2024/07/25 · openalex created_date 2024/07/27 · openalex updated_date 2026/07/22
Abstract Direct RNA sequencing (DRS) is a nanopore-based technique for analyzing RNA in its native form, promising 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 N 6 -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, the first comprehensive benchmark of human peripheral blood and quality assessments of RNA therapeutics.