Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP) is a fluorescent reporter gene commonly used in molecular biology research. Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP) is produced through in vitro transcription. By simulating the mRNA processing process in eukaryotes, it has a 5 'end Cap 1 cap structure, a 3' end poly (A) tail, Cy5-UTP modification, and N1-Me-pUTP modification (Cy5-UTP: N1-Me-pUTP=3:1 (molar ratio)), which increases the stability and translation efficiency of mRNA[1]. Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP) can be used as a standard to detect the transfection efficiency of different transfection reagents, and can also be used as a control to study the transfection and expression of fluorescent proteins in mammalian cells.
N1-Me-pUTP is a methyl modification of naturally occurring pseudouridine pUTP, catalyzed by N1 specific pseudouridine methyltransferase Nepl present in archaea and eukaryotes[2]. This product uses N1-Me-pUTP instead of UTP, effectively enhancing RNA stability while reducing anti RNA immune response[3].
Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP) can directly express proteins in the cytoplasm without relying on promoters, with a faster protein expression rate than transfected DNA. The protein expression level is directly related to the mRNA transfection level, and there is no risk of gene integration. After transfecting cells with N1-Me-pUTP (5'CAP), Cy5 mCherry mRNA can express a strong and bright red fluorescent protein mCherry, with excitation/emission wavelengths of 587/610nm, respectively. Cy5 is a commonly used cyanine fluorescent dye, with maximum excitation/emission wavelengths of 650/670nm, which can monitor the transfection, localization, and expression of the target protein in cells in real time.
References:
[1]. Jemielity J, Fowler T, Zuberek J, et al. Novel "anti-reverse" cap analogs with superior translational properties. RNA. 2003;9(9):1108-1122.
[2]. Callum J C Parr, et al.N 1-Methylpseudouridine substitution enhances the performance of synthetic mRNA switches in cells. 2020 Apr 6;48(6):e35. doi: 10.1093/nar/gkaa070.
[3]. Pedro Morais, Hironori Adachi, Yi-Tao Yu.The Critical Contribution of Pseudouridine to mRNA COVID-19 Vaccines. 2021 Nov 4;9:789427. doi: 10.3389/fcell.2021.789427.
Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP)是一种常用于分子生物学研究的荧光报告基因。Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP)是通过体外转录产生的，通过模拟真核生物中mRNA加工过程具有5'端Cap 1帽结构、3'端poly(A)尾、Cy5-UTP修饰以及N1-Me-pUTP修饰（Cy5-UTP：N1-Me-pUTP=3:1（摩尔比）），增加了mRNA的稳定性和翻译效率[1]。Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP)能够作为标准品检测不同转染试剂的转染效率，也能够作为对照研究哺乳动物细胞中荧光蛋白的转染和表达。
N1-Me-pUTP是天然存在的假尿苷pUTP的甲基修饰物，由存在于古细菌和真核生物中的N1特异性假尿苷甲基转移酶Nepl催化生成[2]。本产品使用N1-Me-pUTP替代UTP，有效增强了RNA稳定性，同时降低抗RNA免疫应答[3]。
Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP)能够在不依赖于启动子的情况下直接在细胞质中表达蛋白，蛋白表达速度比转染DNA更快，蛋白表达量与mRNA的转染量直接相关，并且没有基因整合的风险。Cy5 mCherry mRNA with N1-Me-pUTP (5'CAP)转染细胞后能够表达强烈且明亮的红色荧光蛋白mCherry，激发/发射光波长分别为587/610nm。Cy5是一种常用的花青类荧光染料，最大激发/发射光波长分别为650/670nm，能够实时监测细胞中目的蛋白的转染、定位和表达情况。