N1-methylpseudouridine found within COVID-19 mRNA vaccines produces faithful protein products

• DOI: 10.1016/j.celrep.2022.111300
• Publication Date: 2022-08-15T07:54:18Z
• Authors: Kyusik Q. Kim; Bhagyashri D. Bur...; Shin-Cheng Tzeng; Crystal Jing; Courtney Jungers; Junya Zhang; Liewei L. Yan; Richard D. Vierstra; Sergej Djuranovic; Bradley S. Evans; Hani S. Zaher
• Keywords: 0301 basic medicine; Vaccines; Vaccines, Synthetic; COVID-19 Vaccines; Nucleotides; Messenger; Synthetic; ICTS (Institute of Clinical and Translational Sciences); COVID-19; Proteins; Article; 3. Good health; 03 medical and health sciences; Medicine and Health Sciences; Humans; RNA; RNA, Messenger; mRNA Vaccines; Pseudouridine

Synthetic mRNA technology is a promising avenue for treating and preventing disease. Key to the technology is the incorporation of modified nucleotides such as N1-methylpseudouridine (m1Ψ) to decrease immunogenicity of the RNA. However, relatively few studies have addressed the effects of modified nucleotides on the decoding process. Here, we investigate the effect of m1Ψ and the related modification pseudouridine (Ψ) on translation. In a reconstituted system, we find that m1Ψ does not significantly alter decoding accuracy. More importantly, we do not detect an increase in miscoded peptides when mRNA containing m1Ψ is translated in cell culture, compared with unmodified mRNA. We also find that m1Ψ does not stabilize mismatched RNA-duplex formation and only marginally promotes errors during reverse transcription. Overall, our results suggest that m1Ψ does not significantly impact translational fidelity, a welcome sign for future RNA therapeutics.