Document Type
Article
Publication Date
6-11-2024
Abstract
Nucleoside-modified messenger RNA (mRNA) technologies necessarily incorporate N1-methylpseudouridine into the mRNA molecules to prevent the over-stimulation of cytoplasmic RNA sensors. Despite this modification, mRNA concentrations remain mostly determined through the measurement of UV absorbance at 260 nm wavelength (A260). Herein, we report that the N1-methylpseudouridine absorbs approximately 40% less UV light at 260 nm than uridine, and its incorporation into mRNAs leads to the under-estimation of nucleoside-modified mRNA concentrations, with 5%–15% error, in an mRNA-sequence-dependent manner. We therefore examined the RNA quantification methods and developed the mRNACalc webserver. It accounts for the molar absorption coefficient of modified nucleotides at 260 nm wavelength, the RNA composition of the mRNA, and the A260 of the mRNA sample to enable accurate quantification of nucleoside-modified mRNAs.
Keywords
modified-nucleoside, mRNA, MT: bioinformatics, N1-methylpseudouridine, pseudouridine, UV absorption
Language
English
Publication Title
Molecular Therapy Nucleic Acids
Grant
2246805
Rights
© 2024 The Author(s). This is an Open Access work distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Recommended Citation
Esteban Finol, Sarah E. Krul, Sean J. Hoehn, Xudong Lyu, Carlos E. Crespo-Hernández, The mRNACalc webserver accounts for the N1-methylpseudouridine hypochromicity to enable precise nucleoside-modified mRNA quantification, Molecular Therapy - Nucleic Acids, Volume 35, Issue 2, 2024, 102171, ISSN 2162-2531, https://doi.org/10.1016/j.omtn.2024.102171.
Manuscript Version
Final Publisher Version