National Key Laboratory of Immunity and Inflammation, and CAMS Key Laboratory of Synthetic Biology Regulatory Elements, Suzhou Institute of Systems Medicine, Chinese Academy of Medical Sciences & Peking Union Medical College, Suzhou, China.
Methods Mol Biol. 2025;2854:127-141. doi: 10.1007/978-1-0716-4108-8_14.
Luciferase reporter systems are commonly used in scientific research to investigate a variety of biological processes, including antiviral innate immunity. These systems employ the use of luciferase enzymes derived from organisms such as fireflies or renilla reniformis, which emit light upon reaction with a substrate. In the context of antiviral innate immunity, the luciferase reporter systems offer a noninvasive and highly sensitive approach for real-time monitoring of immune responses in vitro and in vivo, enabling researchers to delve into the intricate interactions and signaling pathways involved in host-virus dynamic interactions. Here, we describe the methods of the promoter-luciferase reporter and enhancer-luciferase reporter, which provide insights into the transcriptional and post-transcriptional regulation of antiviral innate immunity. Additionally, we outline the split-luciferase complementary reporter method, which was designed to explore protein-protein interactions associated with antiviral immunity. These methodologies offer invaluable knowledge regarding the molecular mechanisms underlying antiviral immune pathways and have the potential to support the development of effective antiviral therapies.
荧光素酶报告系统常用于科学研究中，以研究各种生物过程，包括抗病毒固有免疫。这些系统采用来自萤火虫或肾形海肾等生物体的荧光素酶酶，在与底物反应时会发出光。在抗病毒固有免疫方面，荧光素酶报告系统为体外和体内实时监测免疫反应提供了一种非侵入性和高灵敏度的方法，使研究人员能够深入研究宿主-病毒动态相互作用中涉及的复杂相互作用和信号通路。在这里，我们描述了启动子-荧光素酶报告和增强子-荧光素酶报告方法，这些方法提供了对抗病毒固有免疫转录和转录后调控的深入了解。此外，我们还概述了设计用于探索与抗病毒免疫相关的蛋白-蛋白相互作用的分裂荧光素酶互补报告方法。这些方法为抗病毒免疫途径的分子机制提供了宝贵的知识，并有可能支持有效的抗病毒治疗方法的开发。
