Cy5 OVA mRNA with N1-Me-pUTP (5'CAP) is produced through in vitro transcription. By simulating the mRNA processing 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].
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 is a commonly used cyanine fluorescent dye with maximum excitation/emission wavelengths of 650/670nm, capable of real-time monitoring of the transfection, localization, and expression of target proteins in cells.
Ovalbumin (OVA) is a member of the chromoprotein superfamily and the main protein component in egg white. OVA is a glycoprotein with a molecular weight of approximately 45000 daltons that can induce moderate immunity in the body and is a commonly used antigen in immune and biochemical research. Egg white protein is often used as an immunogen for immune experiments, such as establishing animal models of high altitude sickness, asthma, etc. Cy5 OVA 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.
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 OVA 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]。
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。
卵白蛋白(Ovalbumin，OVA)是色蛋白超家族的成员，也是蛋清中的主要蛋白成分。OVA是一种分子量约45000道尔顿的糖蛋白，能够引起机体适度的免疫性，是免疫和生化研究的常用抗原。卵白蛋白常被用作免疫原进行免疫实验，如高原反应性、哮喘等动物模型建立。Cy5 OVA 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。