Fluc-eGFP fluorescent protein is a fluorescent reporter gene commonly used in molecular biology research. This product connects firefly luciferase mRNA and green fluorescent protein EGFP mRNA through a Linker, and can be used for the detection of two reporter gene experiments. Fluc-eGFP mRNA can express protein directly in the cytoplasm without relying on a promoter. The protein expression speed is faster than transfection of deoxyribonucleotides. The protein expression amount is directly related to the transfection amount of mRNA, and there is no gene Integration Risks. Fluc-eGFP mRNA transfected cells can express strong and bright green fluorescent protein eGFP and firefly luciferase protein. The excitation/emission wavelengths of eGFP are 488 nm/509 nm respectively; firefly luciferase catalyzes luciferin or fatty aldehydes in organisms to produce autofluorescence and chemiluminescence, with wavelengths of approximately 550-570nm[1].
By simulating the mRNA processing process in eukaryotes, the product has a Cap 1 cap structure at the 5' end and a poly(A) tail at the 3' end, which increases the stability and translation efficiency of the mRNA[2]. N1-Me-pUTP is a methyl modification of the naturally occurring pseudouridine pUTP, which is catalyzed by the N1-specific pseudouridine methyltransferase Nepl that exists in archaea and eukaryotes[3]. This product uses N1-Me-pUTP instead of UTP, which effectively enhances RNA stability and reduces anti-RNA immune responses[4].
References:
[1]. JoÃo M M LeitÃo, Joaquim C G Esteves da Silva. Firefly luciferase inhibition. 2010 Oct 5;101(1):1-8. doi: 10.1016/j.jphotobiol.2010.06.015. Epub 2010 Jul 3.
[2]. Jemielity J, Fowler T, Zuberek J, et al. Novel "anti-reverse" cap analogs with superior translational properties. RNA. 2003;9(9):1108-1122
[3]. 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.
[4]. 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.
Fluc-eGFP荧光蛋白是一种常用于分子生物学研究的荧光报告基因。该产品通过Linker连接萤火虫荧光素酶mRNA和绿色荧光蛋白EGFP mRNA,可用于两种报告基因实验的检测。Fluc-eGFP mRNA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脱氧核糖核苷酸更快,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。Fluc-eGFP mRNA转染细胞后能够表达强烈且明亮的绿色荧光蛋白eGFP和萤火虫荧光素酶蛋白。eGFP激发/发射光波长分别为488 nm/509 nm；萤火虫荧光素酶催化生物体中的荧光素或脂肪醛产生自发荧光和化学发光,波长约为550- 570nm[1]。
通过模拟真核生物中mRNA加工过程,该产品的5'端具有Cap 1帽结构,3'端具有poly(A)尾,增加了mRNA的稳定性和翻译效率[2]。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[3]。该产品使用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[4]。