N 1 -methylpseudouridylation of mRNA causes +1 ribosomal frameshifting.

Autor: Mulroney TE; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Pöyry T; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Yam-Puc JC; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Rust M; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Harvey RF; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Kalmar L; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Horner E; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Booth L; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Ferreira AP; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Stoneley M; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Sawarkar R; MRC Toxicology Unit, University of Cambridge, Cambridge, UK., Mentzer AJ; Wellcome Centre for Human Genetics, University of Oxford, Oxford, UK., Lilley KS; Department of Biochemistry, University of Cambridge, Cambridge, UK., Smales CM; School of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, UK.; National Institute for Bioprocessing Research and Training, University College Dublin, Foster Avenue, Mount Merrion, Dublin, Ireland., von der Haar T; School of Biosciences, Division of Natural Sciences, University of Kent, Canterbury, UK., Turtle L; NIHR Health Protection Research Unit for Emerging and Zoonotic Infections, Institute of Infection, Veterinary and Ecological Sciences, University of Liverpool, Liverpool, UK., Dunachie S; NIHR Oxford Biomedical Research Centre, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.; NDM Centre for Global Health Research, Nuffield Department of Medicine, University of Oxford, Oxford, UK.; Mahidol-Oxford Tropical Medicine Research Unit, Mahidol University, Bangkok, Thailand., Klenerman P; NIHR Oxford Biomedical Research Centre, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.; Translational Gastroenterology Unit, Nuffield Department of Medicine, University of Oxford, Oxford, UK., Thaventhiran JED; MRC Toxicology Unit, University of Cambridge, Cambridge, UK. jedt2@cam.ac.uk., Willis AE; MRC Toxicology Unit, University of Cambridge, Cambridge, UK. aew80@cam.ac.uk.
Jazyk: angličtina
Zdroj: Nature [Nature] 2024 Jan; Vol. 625 (7993), pp. 189-194. Date of Electronic Publication: 2023 Dec 06.
DOI: 10.1038/s41586-023-06800-3
Abstrakt: 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 effect 1,2 . Modified ribonucleotides are commonly incorporated into therapeutic IVT mRNAs to decrease their innate immunogenicity 3-5 , 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.
(© 2023. The Author(s).)
Databáze: MEDLINE
Externí odkaz: