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In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
TSA (Tyramide Signal Amplification), used for signal amplification of ISH, IHC and IC etc.
Separation of phosphorylated and non-phosphorylated proteins without phospho-specific antibody
A convenient and sensitive way for cell proliferation assay and cytotoxicity assay
Protect the integrity of proteins from multiple proteases and phosphatases for different applications.
N1-Methylpseudouridine is a modified nucleoside used for enhancing mRNA translation. Incorporation of N1-Methylpseudouridine into mRNA turns off the immune/eIF2α phosphorylation-dependent inhibition of translation, and increases ribosome pausing and density on the mRNA, thus dramatically facilitating the translation process. N1-Methylpseudouridine outperforms several other modified nucleosides (e.g. 5-Methylcytidine) and corresponding combinations in terms of translation capacity.
References:
1. Svitkin YV, Cheng YM, Chakraborty T, et al. N1-methyl-pseudouridine in mRNA enhances translation through eIF2α-dependent and independent mechanisms by increasing ribosome density. Nucleic Acids Research, 2017, 45(10): 6023-6036.
2. Andries O, Mc Cafferty S, De Smedt SC, et al N(1)-methylpseudouridine-incorporated mRNA outperforms pseudouridine-incorporated mRNA by providing enhanced protein expression and reduced immunogenicity in mammalian cell lines and mice. Journal of Controlled Release, 2015, 217: 337-344.
Cell lines
Various mammalian cells including A549, BJ, C2C12, HeLa and primary keratinocytes
Reaction Conditions
24 h incubation
Applications
In various mammalian cells, mRNAs in the N1-Methylpseudouridine + 5-Methylcytidine group showed reduced cytotoxicity compared to the Pseudouridine + 5-Methylcytidine group. The mRNAs simultaneously modified by N1-Methylpseudouridine and 5-Methylcytidine also exhibited reduced activation of the intracellular innate immune response.
Animal models
7-week-old Balb/c mice
Dosage form
20 μg
Once daily through intradermal or intramuscular routes by lipofection, for 21 days
N1-Methylpseudouridine alone and its combination with 5-Methylcytidine outperformed the current state-of-the-art Pseudouridine and its combination with 5-Methylcytidine, in terms of translation capacity.
Note
The technical data provided above is for reference only.