Luciferase(Fluc)can detect gene expression extremely sensitively and efficiently, so it is often used as a bioluminescent reporter gene for gene regulation and functional studies. Fluc mRNA can express proteins directly in the cytoplasm without relying on a promoter. The protein expression speed of Fluc mRNA is faster than transfected DNA. The protein expression level is directly related to the transfection amount of mRNA, and there is no risk of gene integration. Luciferase protein catalyzes intracellular luciferin or fatty aldehyde to produce autofluorescence, producing chemiluminescence at a wavelength of about 550-570nm[1].
Luciferase can be used to detect promoter activity or dual fluorescent molecule complementation experiments. Firefly luciferase and Renilla luciferase can respectively catalyze their respective substrates to emit fluorescence of different colors. The two light absorption wavelengths are different and the detection does not interfere with each other. Therefore, they can be used as dual luciferases in the same chemical reaction system. Reporter gene systems are used simultaneously[2].
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[3].
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]. Yong Zhong Xu, et. Promoter deletion analysis using a dual-luciferase reporter system.2013;977:79-93. doi: 10.1007/978-1-62703-284-1_7.
[3]. Jemielity J, Fowler T, Zuberek J, et al. Novel "anti-reverse" cap analogs with superior translational properties. RNA. 2003;9(9):1108-1122