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  • 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
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glycerol, for molecular biology, ≥99.0%
Sigma-Aldrich
TWEEN® 20, viscous liquid
Sigma-Aldrich
Interferon-γ human, 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)