Institute of Molecular Virology, Ulm University Medical Center, 89081, Ulm, Baden-Württemberg, Germany.
STAR Protoc. 2021 Dec 17;2(4):100781. doi: 10.1016/j.xpro.2021.100781. Epub 2021 Aug 13.
We present a protocol for analyzing the impact of SARS-CoV-2 proteins in interferon signaling using luciferase reporter assays. Here, the induction of defined promoters can be quantitatively assessed with high sensitivity and broad linear range. The results are similar to those obtained using qPCR to measure endogenous mRNA induction. The assay requires stringent normalization and confirmation of the results in more physiological settings. The protocol is adaptable for other viruses and other innate immune stimuli. For complete details on the use and execution of this protocol, please refer to Hayn et al. (2021).
我们提出了一种使用荧光素酶报告基因检测分析 SARS-CoV-2 蛋白对干扰素信号影响的方案。在该方案中,可以用高灵敏度和宽线性范围定量评估特定启动子的诱导。其结果与使用 qPCR 测量内源性 mRNA 诱导的结果相似。该检测需要严格的归一化,并在更接近生理的条件下对结果进行确认。该方案可适用于其他病毒和其他先天免疫刺激物。如需了解该方案的使用和实施的详细信息,请参考 Hayn 等人(2021 年)的研究。