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Merck
  • Luciferase reporter assays to monitor interferon signaling modulation by SARS-CoV-2 proteins.

Luciferase reporter assays to monitor interferon signaling modulation by SARS-CoV-2 proteins.

STAR protocols (2021-08-19)
Maximilian Hirschenberger, Manuel Hayn, Alexandre Laliberté, Lennart Koepke, Frank Kirchhoff, Konstantin Maria Johannes Sparrer
摘要

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).

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Sigma-Aldrich
甘油, for molecular biology, ≥99.0%
Sigma-Aldrich
TWEEN ® 20, viscous liquid
Sigma-Aldrich
干扰素β蛋白,重组人类, Recombinant Human Interferon Beta 1a (Hu-IFNbeta 1a).
Sigma-Aldrich
干扰素-γ 人, recombinant, expressed in E. coli, ≥98% (SDS-PAGE), lyophilized powder, suitable for cell culture
Sigma-Aldrich
IFN-alpha 2a human, recombinant, expressed in E. coli, ≥95% (SDS-PAGE), ≥95% (HPLC)