N1-Methylpseudo-UTP

Product Information

N1-Methylpseudo-UTP – A Nucleotide for the Synthesis of Immunosilent mRNA Through In Vitro Transcription

N1-methylpseudouridine 5´-triphosphate (m¹ΨTP) is a modified ribonucleotide analog widely used in in vitro transcription (IVT) for the synthesis of mRNA to enhance biological stability and reduced immunogenicity.^[1] When incorporated into mRNA transcripts, m¹Ψ improves translational efficiency and strongly suppresses activation of innate immune sensors such as TLRs and RIG‑I pathways.

These properties make m¹ΨTP a key component in the development of therapeutic mRNA vaccines, protein‑replacement therapies, and gene‑modulation technologies.^[2]

Applications of N1‑Methylpseudouridine

m¹Ψ is widely used in biotechnology, vaccine development, and therapeutic mRNA manufacturing. Substituting UTP with m¹ΨTP during IVT yields mRNA with:

• Reduced innate immune activation (immunosilent mRNA)
• Increased mRNA stability
• Higher translational efficiency
• Improved protein expression in vivo and in vitro

These advantages were critical in the success of mRNA‑based COVID‑19 vaccines and are now standard in next‑generation mRNA therapeutics.

m¹Ψ‑modified mRNA is used for:

• mRNA vaccine development
• Therapeutic mRNA research
• Functional protein expression studies
• Gene modulation and gene editing approaches
• Development of LNP‑formulated mRNA delivery systems

Because m¹ΨTP can directly replace UTP in IVT reactions (1:1), it is simple to integrate into existing mRNA workflows.

baseclick supplies high‑purity, reliable m¹ΨTP optimized for use in mRNA research:

• Market‑leading pricing with attractive bulk options
• ≥ 98.5 % HPLC purity for consistent, high performance IVT reactions
• Available Sizes: 1 µmol, 5 µmol, and bulk quantities upon request (research use only).
• Fast, reliable shipping for research and development needs

IVT Kits Containing N1‑Methylpseudouridine

baseclick provides an IVT solution incorporating m¹ΨTP:

• baseScribe Mod‑Methylpseudo Cap1 Kit

This kit enables production of up to 160 µg of CleanCap (Cap1)‑capped, m¹Ψ‑modified mRNA per 20 µL reaction from DNA templates with a T7 promoter. The resulting mRNA contains 100% uridine replacement by m¹Ψ, ensuring high expression and minimized immunogenicity for both in vitro and in vivo applications.

Ready‑to‑Use N1‑Methylpseudouridine‑Modified mRNA from baseclick

Through our partnership with Eurofins Genomics, baseclick offers fully customized, ready‑to‑use mRNA from basic expression constructs to highly functionalized mRNA for advanced applications.

Our joint mRNA service includes:

•  Plasmid DNA production (via Eurofins)
• High-yield IVT mRNA synthesis (via baseclick)
• 5′-Capping & poly(A) tailing
• Internal base modifications (e.g., N1-methylpseudouridine, 5-ethynyl uridine)
• Site-specific click conjugation (e.g. with fluorophores, biotin, or other ligands)
• Quality control and data documentation
• Support for downstream LNP encapsulation or in vivo delivery

Biological Function of N1‑Methylpseudouridine

m¹Ψ is essential for generating immunosilent, highly translated mRNA.^[2] It reduces activation of innate immune pathways and significantly enhances protein yield, properties that were instrumental in the success of BioNTech/Pfizer’s Comirnaty COVID‑19 vaccine.^[3]

The discovery of pseudouridine and N1‑methylpseudouridine as immune‑silent mRNA nucleotides by Karikó and Weissman revolutionized mRNA therapeutics and was awarded the 2023 Nobel Prize in Physiology or Medicine.^[1]

m¹ΨTP can seamlessly replace UTP in IVT reactions, making it the preferred uridine analog in modern mRNA technology.

LITERATURE

[1] Incorporation of Pseudouridine Into mRNA Yields Superior Nonimmunogenic Vector With Increased Translational Capacity and Biological Stability. K. Karikó, H. Muramatsu, F. A Welsh, J. Ludwig, H. Kato, S. Akira, D. Weissman, 2008, Mol. Ther., Vol. 16(11), p. 1833–1840.

[2] mRNA-based therapeutics — developing a new class of drugs. U. Sahin, K. Karikó, Ö. Türeci, 2014, Nat. Rev. Drug Discov., Vol. 13, p. 759–780.

[3] Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. F. P. Polack et al. for the C4591001 Clinical Trial Group, 2020, N. Engl. J. Med., Vol. 383(27), p. 2603–2615.

N1-methylpseudouridine-incorporated mRNA outperforms pseudouridine-incorporated mRNA by providing enhanced protein expression and reduced immunogenicity in mammalian cell lines and mice, O. Andries et al., 2015, Journal of Controlled Release, Vol. 217, p. 337-344.

https://doi.org/10.1016/j.jconrel.2015.08.051

Multicomponent Lipid Nanoparticles for RNA Transfection, N. Gretskaya et al., 2023, Pharmaceutics, Vol. 15(4), 1289.

https://doi.org/10.3390/pharmaceutics15041289

CpG oligodeoxynucleotide developed to activate primate immune responses promotes antitumoral effects in combination with a neoantigen-based mRNA cancer vaccine, Q. Li et al., 2021, Drug Design, Development and Therapy, p. 3953-3963.

https://doi.org/10.2147/DDDT.S325790