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Sensing of cytoplasmic DNA by cGAS is essential for the initiation of immune responses against several viruses. cGAS also plays important roles in some autoinflammatory and autoimmune diseases and may be involved in immune responses targeting cancer cells. Once activated, cGAS catalyzes the formation of the di-nucleotide 2'-3'-cyclic GMP-AMP (cGAMP), which propagates a signaling cascade leading to the production of type I interferons (IFNs). Interestingly, cGAMP is incorporated into enveloped viruses and is transferred to newly infected cells by virions. In this article, we describe a method to purify cGAMP from viral particles and a bioassay to measure its activity. This assay takes advantage of a reporter cell line that expresses the genes encoding green fluorescent protein (GFP) and firefly luciferase under the control of the IFNß promoter, allowing the testing of several samples in a single experiment taking not more than 3 days.

Original publication

DOI

10.1007/978-1-4939-7237-1_8

Type

Journal article

Journal

Methods Mol Biol

Publication Date

2017

Volume

1656

Pages

143 - 152

Keywords

Bioassay, Innate immunity, STING, Type I IFN, cGAMP, Humans, Immunity, Innate, Interferon-beta, Nucleotides, Cyclic, THP-1 Cells, Virion, Virus Diseases, Viruses