N1-Me-pUTP,100mM Tris Solution (N1-methyl-pseudouridine-5′-triphosphate) is a modified nucleoside triphosphate, often used as a substitute for UTP in in vitro transcription (IVT). Nucleotide modification can greatly enhance the performance of mRNA by reducing immunogenicity and increasing the stability of RNA molecules[1]. In addition to shutting down immune/eIF2α phosphorylation-dependent translational repression, the incorporated N1-Me-pUTP also significantly altered the dynamics of translation by increasing ribosome pauses and density on mRNA, thereby enhancing mRNA protein expression levels[2]. N1-Me-pUTP, which can greatly enhance the performance of miRNA- and RBP-responsive mRNA switches. The higher sensitivity of N1-Me-pUTP-containing switches for miRNA and RBP (U1A and MS2) allows clear separation of positive and negative miRNA- or RBP-expressing cells[3].
References:
[1] Andries O, Mc Cafferty S, De Smedt SC, Weiss R, Sanders NN, Kitada T. N(1)-methylpseudouridine-incorporated mRNA outperforms pseudouridine-incorporated mRNA by providing enhanced protein expression and reduced immunogenicity in mammalian cell lines and mice. J Control Release. 2015;217:337-344.
[2] Svitkin YV, Cheng YM, Chakraborty T, Presnyak V, John M, Sonenberg N. N1-methyl-pseudouridine in mRNA enhances translation through eIF2α-dependent and independent mechanisms by increasing ribosome density. Nucleic Acids Res. 2017;45(10):6023-6036.
[3] Parr CJC, Wada S, Kotake K, et al. N 1-Methylpseudouridine substitution enhances the performance of synthetic mRNA switches in cells. Nucleic Acids Res. 2020;48(6):e35.
N1-Me-pUTP,100mM Tris Solution是一种修饰的三磷酸核苷酸，通常作为UTP的替代品用于体外转录（IVT）。核苷酸修饰能够通过降低免疫原性和提高RNA分子的稳定性，极大增强mRNA的性能[1]。除了关闭免疫/eIF2α磷酸化依赖性的翻译抑制外，N1-Me-pUTP还通过增加mRNA上的核糖体暂停和密度来显著改变翻译过程的动力学，提高mRNA蛋白表达水平[2]。N1-Me-pUTP能够显著增强miRNA和RBP响应型mRNA开关的性能。含有N1-Me-pUTP的开关对miRNA和RBP（U1A和MS2）具有更高的灵敏度，能够清晰地区分表达miRNA或RBP的阳性细胞和阴性细胞[3]。