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

STAR Protoc. 2021 Dec 17;2(4):100781. doi: 10.1016/j.xpro.2021.100781. Epub 2021 Aug 13.

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).

Keywords: Cell-based assays; Gene expression; Immunology; Microbiology; Molecular biology; Molecular/chemical probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / pharmacology
  • COVID-19 / metabolism
  • COVID-19 / pathology*
  • COVID-19 / virology
  • COVID-19 Drug Treatment
  • Gene Expression Regulation, Viral / drug effects*
  • Humans
  • Interferons / pharmacology*
  • Luciferases / genetics
  • Luciferases / metabolism*
  • Promoter Regions, Genetic
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • SARS-CoV-2 / drug effects
  • SARS-CoV-2 / metabolism*
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • Antiviral Agents
  • RNA, Messenger
  • Viral Proteins
  • Interferons
  • Luciferases