| Chapter title |
Purification of Cyclic GMP-AMP from Viruses and Measurement of Its Activity in Cell Culture
|
|---|---|
| Chapter number | 8 |
| Book title |
Innate Antiviral Immunity
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-7237-1_8 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7236-4, 978-1-4939-7237-1
|
| Authors |
Alice Mayer, Jonathan Maelfait, Anne Bridgeman, Jan Rehwinkel, Mayer, Alice, Maelfait, Jonathan, Bridgeman, Anne, Rehwinkel, Jan |
| Abstract |
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. |
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| Unknown | 1 | 25% |
Demographic breakdown
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Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 27% |
| Other | 2 | 13% |
| Student > Bachelor | 2 | 13% |
| Researcher | 2 | 13% |
| Student > Master | 1 | 7% |
| Other | 1 | 7% |
| Unknown | 3 | 20% |
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|---|---|---|
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| Agricultural and Biological Sciences | 1 | 7% |
| Neuroscience | 1 | 7% |
| Other | 1 | 7% |
| Unknown | 3 | 20% |