Biomedical Research Institute, AIST, Tsukuba, Ibaraki 305-8566, Japan.
Department of Biomedical Engineering, Osaka Institute of Technology (OIT), Osaka, Osaka 535-8585, Japan.
Int J Mol Sci. 2020 Dec 24;22(1):119. doi: 10.3390/ijms22010119.
Both fluorescent and luminescent observation are widely used to examine real-time gene expression patterns in living organisms. Several fluuorescent and luminescent proteins with specific optical properties have been developed and applied for simultaneous, multi-color observation of more than two gene expression profiles. Compared to fluorescent proteins, however, the application of multi-color luminescent imaging in living organisms is still limited. In this study, we introduced two-color luciferases into the soil nematode and performed simultaneous analysis of two gene expression profiles. Using a green-emitting luciferase Eluc (emerald luciferase) and red-emitting luciferase SLR (stable luciferase red), the expression patterns of two genes were simultaneously observed in single animals from embryonic to adult stages over its whole life span. In addition, dual gene activities were observed at the single embryo level, with the simultaneous observation of morphological changes. These are the first application of a two-color luciferase system into a whole animal and suggest that precise relationship of expression patterns of multiple genes of interest can be analyzed over the whole life of the animal, dependent on the changes in genetic and/or environmental conditions.
荧光和发光观察广泛用于研究生物体内实时基因表达模式。已经开发和应用了几种具有特定光学特性的荧光和发光蛋白，用于同时观察两种以上基因表达谱的多色观察。然而，与荧光蛋白相比，多色发光成像在生物体内的应用仍然有限。在这项研究中，我们将双色荧光素酶引入土壤线虫中，并进行了两种基因表达谱的同时分析。使用绿色发光荧光素酶 Eluc（祖母绿荧光素酶）和红色发光荧光素酶 SLR（稳定红色荧光素酶），在整个生命周期内，从胚胎到成年阶段，在单个动物中同时观察两种基因的表达模式。此外，在单个胚胎水平上观察到双重基因活性，并同时观察形态变化。这是首次将双色荧光素酶系统应用于整个动物，并表明可以根据遗传和/或环境条件的变化，在动物的整个生命周期内分析多个感兴趣基因表达模式的精确关系。
