Incorporation of pseudouridine into mRNA yields superior nonimmunogenic vector with increased translational capacity and biological stability.

By K. Karikó, Hiromi Muramatsu, F. Welsh, J. Ludwig, H. Kato, Shizuo Akira, D. Weissman. 2008

1,650 citations recorded by the source corpus.

In vitro-transcribed mRNAs encoding physiologically important proteins have considerable potential for therapeutic applications. However, in its present form, mRNA is unfeasible for clinical use because of its labile and immunogenic nature. Here, we investigated whether incorporation of naturally modified nucleotides into transcripts would confer enhanced biological properties to mRNA. We found that mRNAs containing pseudouridines have a higher translational capacity than unmodified mRNAs when tested in mammalian cells and lysates or administered intravenously into mice at 0.015-0.15 mg/kg doses. The delivered mRNA and the encoded protein could be detected in the spleen at 1, 4, and 24 hours after the injection, where both products were at significantly higher levels when pseudouridine-containing mRNA was administered. Even at higher doses, only the unmodified mRNA was immunogenic, inducing high serum levels of interferon-alpha (IFN-alpha). These findings indicate that nucleoside modification is an effective approach to enhance stability and translational capacity of mRNA while diminishing its immunogenicity in vivo. Improved properties conferred by pseudouridine make such mRNA a promising tool for both gene replacement and vaccination.

Subfields: Immunology, Molecular Biology

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Citations in the corpus

mRNA Vaccines in the COVID-19 Pandemic and Beyond. (2021, background): “In contrast, incorporation of modified nucleosides such as pseudouridine was shown to reduce or abrogate this innate sensing (60–62), allowing a high level of protein expression in vivo (63, 64).”
Generating the optimal mRNA for therapy: HPLC purification eliminates immune activation and improves translation of nucleoside-modified, protein-encoding mRNA (2011, methodology): “Human and murine dendritic cells (DCs) were generated as described (5).”
mRNA-based therapeutics: powerful and versatile tools to combat diseases (2022, background): “Previous research has indicated that nucleoside modifications improved mRNA translation efficiency (Ψ, 5mC, Ψ/5mC or N1-methyl-pseudouridine/5methylcytidine), and the pseudouridine/5-methylcytidine (Ψ/ 5mC)-modified mRNA partly suppressed the innate immune activation by mRNA vaccines and increased t”
Incorporation of pseudouridine into mRNA enhances translation by diminishing PKR activation (2010, methodology): “We previously reported that mRNA transcribed in vitro containing in place of uridine is translated more efficiently than mRNA containing unmodified nucleosides (33).”
mRNA-based therapeutics — developing a new class of drugs (2014, positive): “…naturally occurring modified nucleosides such as pseudouridine, 2-thiouridine, 5-methyluridine, 5-methylcytidine or N6-methyladenosine into the IVT mRNA; this has been shown to suppress both the intrinsic adjuvant activity of IVT mRNA as well as its inhibitory effects on translation 75,245 .”