N1-methylpseudo-UTP is a nucleobase-modified nucleotide, used for synthesizing mRNA with reduced immunogenicity and improved stability. It was reported that N1-methylpseudouridine (m1Ψ) modification alone or in combination with 5-methylcytidine (m5C) exhibited superiority over the current state-of-the-art pseudouridine (Ψ) or m5C/Ψ-modified mRNA platform by providing up to ~ 44-fold (when comparing double modified mRNAs) and ~ 13-fold (when comparing single modified mRNAs) higher reporter gene expression in cells and mice, respectively. Moreover, compared with (m5C/)Ψ-modified mRNAs, (m5C/)m1Ψ-modified mRNAs showed reduced intracellular innate immunogenicity and resulted in improved cellular viability after in vitro transfection. Thus, N1-methylpseudo-UTP might serve as a useful ingredient for synthesizing drugable mRNAs with better performance.
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