2026-10-14T17:54:17Z https://api.figshare.com/v2/oai

oai:figshare.com:article/33120299 2026-09-16T08:55:38Z category_24178 item_type_3 month_year_09_2026

FCS raw data for Poulis, Robecchi et al, Nature Communications, 2026 Giovanni Robecchi (24480158) Panagiotis Poulis (22660170) Michele Felletti (22660173) Arina Kurochkina (22660165) Marina V. Rodnina (21635641) Biochemistry and cell biology not elsewhere classified biochemistry Flow cytometry Frameshifting <p dir="ltr"><i>N</i>^<em>1</em>-methylpseudouridine (m¹Ψ) is a key modification in mRNA vaccines against SARS-CoV-2, introduced to reduce immunogenicity and increase mRNA stability. Given the widespread clinical use, understanding its impact on translation fidelity is essential for vaccine safety. Recent studies suggested that m¹Ψ can promote ribosomal frameshifting, a translational error that generates aberrant peptides. These peptides are presented on major histocompatibility complex and elicit immune responses, raising concerns about unintended antigenicity. To address this, we systematically examined the efficiency of frameshifting induced by m¹Ψ and the underlying mechanism, aiming to inform future mRNA vaccine design. Using mRNA-based dual-fluorescence reporters in cells, fully <i>in-vitro</i> reconstituted translation system,<i> </i>and single-molecule FRET microscopy, we show that m¹Ψ modestly increases +1 frameshifting on UUUC motifs in therapeutic mRNAs. Frameshifting occurs when the peptidyl-tRNA pauses in the P site of the ribosome awaiting delivery of the next aminoacyl-tRNA. m¹Ψ both weakens codon-anticodon interactions and promotes a frameshifting-prone conformation of the P-site peptidyl-tRNA, thereby increasing slippage during the pause. Replacing the slippery UUU C motif with UUC C or UUU U eliminates this effect. Our results reveal how a clinically relevant mRNA modification promotes translational recoding and show that codon optimization can mitigate this risk in therapeutic mRNA design.</p> 2026-09-16T08:55:38Z Dataset Dataset 10.6084/m9.figshare.33120299.v1 https://figshare.com/articles/dataset/FCS_raw_data_for_Poulis_Robecchi_et_al_Nature_Communications_2026/33120299 CC BY 4.0