海肾萤光素酶发光体系简单，不需要ATP、Mg2+等辅助因子，能够表达于哺乳动物细胞中并且无细胞毒性，已经被成功应用于检测活性氧、目标基因的表达、蛋白-蛋白相互作用、药物高通量筛选等。但该生物发光体系还存在发射波长短、发光强度低、容易被组织吸收、作用时间短、稳定性较差等问题，其主要原因是腔肠素类似物不能够很好地被海肾萤光素酶所识别。本项目拟在大量的前期工作基础上，通过对腔肠素与海肾萤光素酶结合模式研究，在海肾萤光素酶底物腔肠素的C-2、C-3、C-6和C-8位合理引入能提高亲和力、改善细胞组织通透性和体内药物代谢动力学性质的杂环基团，建立和开发基于海肾萤光素酶体系的高灵敏的新型生物发光底物并研究其生物发光活性，同时采用此生物发光体系进行报告基因系统的构建，最终形成发光底物改造、细胞活性研究、报告基因系统构建、动物成像分析的腔肠素生物发光体内可视化分析平台。

As one of the common bioluminescence systems, renilla luciferase luminescence system requires renilla luciferase, coelenterazine and oxygen to produce a visible light. It should be noted that renilla luciferase system is too simple to need ATP, Mg2+ and other cofactors. Moreover, renilla luciferase can be expressed in mammalian cells with low cytotoxicity. However, if widely used for animal imaging, such a bioluminescence system is necessary to overcome the following shortcomings, including short emission wavelength, short imaging duration, low sensitivity and stability. In the current project, on the basis of our fruitful preliminary results, the PI aims at obtaining novel substrates with high sensitivity for renilla luciferin-coelenterazine bioluminescence system by rational modifying the chemical structure of coelenterazine and introducing heterocyclic rings with increasing affinity, high premeable and well tolerable ability in vivo at the C-2, C-3, C-6 and C-8 position, and examining these proposed substrates' bioluminescent imaging potential on the molecular, cellular and living animal levels. The successful results from this project will develop a visualization analysis platform of renilla bioluminescence, including development of molecular probe, biomolecule recognition, as well as generation of cell and animal model.