A false sense of circularity: hidden variability in RNase R enzymes confounds reliable circRNA validation
- 1Ghent University
Abstract
Circular RNAs (circRNAs) are an emerging class of RNA molecules involved in development and disease. They are generated through back-splicing from pre-mRNA. RNaseR treatment, which degrades linear RNA while preserving circRNAs, is widely used for circRNA validation and enrichment. This short report demonstrated differential performance of two RNaseR enzymes from different suppliers, despite identical experimental conditions, indicating that these enzymes should not be considered interchangeable across suppliers. Importantly, these findings highlight that without demonstrating efficient linear RNA degradation, unchanged circRNA abundance after RNase R treatment is inherently uninformative and may contribute to false-positive circRNA validation or inadequate circRNA enrichment. We therefore highlight that RNaseR digestion should be consistently verified using a linear control, particularly when changing enzyme suppliers.
Related
- A positive feedback loop of kynurenine-AhR-EMT upregulates tryptophan metabolic enzymes via epigenetic modification to facilitate immune escape in triple-negative breast cancer
- Functional differences of two homologous enzymes for LacdiNAc biosynthesis: B4GALNT3 and B4GALNT4
- Identification of Novel Enzymes Regulating ADP-Ribosylation of Uracil and Thymine in Nucleic Acids
- De novo design of allosterically controlled enzymes
- Harnessing Artificial Intelligence to Understand Enzyme Catalysis and Discover New Enzymes
BibTeX
@misc{ulrix2026false,
title = {{A false sense of circularity: hidden variability in RNase R enzymes confounds reliable circRNA validation}},
author = {L. Ulrix and J. H. De Waele and J. Vandesompele and M. Hofmans and B. De Moerloose and T. Lammens},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.13.757942},
url = {https://www.biorxiv.org/content/10.64898/2026.10.13.757942v1},
note = {CC BY-ND},
}
RIS
TY - UNPB
TI - A false sense of circularity: hidden variability in RNase R enzymes confounds reliable circRNA validation
AU - L. Ulrix
AU - J. H. De Waele
AU - J. Vandesompele
AU - M. Hofmans
AU - B. De Moerloose
AU - T. Lammens
PY - 2026
DA - 2026/10/13
PB - bioRxiv
DO - 10.64898/2026.10.13.757942
UR - https://www.biorxiv.org/content/10.64898/2026.10.13.757942v1
AB - Circular RNAs (circRNAs) are an emerging class of RNA molecules involved in development and disease. They are generated through back-splicing from pre-mRNA. RNaseR treatment, which degrades linear RNA while preserving circRNAs, is widely used for circRNA validation and enrichment. This short report demonstrated differential performance of two RNaseR enzymes from different suppliers, despite identical experimental conditions, indicating that these enzymes should not be considered interchangeable across suppliers. Importantly, these findings highlight that without demonstrating efficient linear RNA degradation, unchanged circRNA abundance after RNase R treatment is inherently uninformative and may contribute to false-positive circRNA validation or inadequate circRNA enrichment. We therefore highlight that RNaseR digestion should be consistently verified using a linear control, particularly when changing enzyme suppliers.
ER -