N <sup>1</sup>-methylpseudouridylation of mRNA causes +1 ribosomal frameshifting
Issued Date
2023-01-01
Resource Type
ISSN
00280836
eISSN
14764687
Scopus ID
2-s2.0-85178897070
Journal Title
Nature
Rights Holder(s)
SCOPUS
Bibliographic Citation
Nature (2023)
Suggested Citation
Mulroney T.E., Pöyry T., Yam-Puc J.C., Rust M., Harvey R.F., Kalmar L., Horner E., Booth L., Ferreira A.P., Stoneley M., Sawarkar R., Mentzer A.J., Lilley K.S., Smales C.M., von der Haar T., Turtle L., Dunachie S., Klenerman P., Thaventhiran J.E.D., Willis A.E. N <sup>1</sup>-methylpseudouridylation of mRNA causes +1 ribosomal frameshifting. Nature (2023). doi:10.1038/s41586-023-06800-3 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/91489
Title
N <sup>1</sup>-methylpseudouridylation of mRNA causes +1 ribosomal frameshifting
Other Contributor(s)
Abstract
In vitro-transcribed (IVT) mRNAs are modalities that can combat human disease, exemplified by their use as vaccines for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). IVT mRNAs are transfected into target cells, where they are translated into recombinant protein, and the biological activity or immunogenicity of the encoded protein exerts an intended therapeutic effect1,2. Modified ribonucleotides are commonly incorporated into therapeutic IVT mRNAs to decrease their innate immunogenicity3–5, but their effects on mRNA translation fidelity have not been fully explored. Here we demonstrate that incorporation of N 1-methylpseudouridine into mRNA results in +1 ribosomal frameshifting in vitro and that cellular immunity in mice and humans to +1 frameshifted products from BNT162b2 vaccine mRNA translation occurs after vaccination. The +1 ribosome frameshifting observed is probably a consequence of N 1-methylpseudouridine-induced ribosome stalling during IVT mRNA translation, with frameshifting occurring at ribosome slippery sequences. However, we demonstrate that synonymous targeting of such slippery sequences provides an effective strategy to reduce the production of frameshifted products. Overall, these data increase our understanding of how modified ribonucleotides affect the fidelity of mRNA translation, and although there are no adverse outcomes reported from mistranslation of mRNA-based SARS-CoV-2 vaccines in humans, these data highlight potential off-target effects for future mRNA-based therapeutics and demonstrate the requirement for sequence optimization.