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Murine cgas ha

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Murine cGAS-HA is a laboratory tool that allows for the expression and detection of the murine cGAS (cyclic GMP-AMP synthase) protein. cGAS is an important sensor of cytosolic DNA and plays a crucial role in the activation of the innate immune response. The Murine cGAS-HA product provides a means to study the function and regulation of this enzyme in a controlled experimental setting.

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Lab products found in correlation

Pmxs ires puro vector, by Cell Biolabs (2 mentions) Plat e cells, by Cell Biolabs (2 mentions)

2 protocols using murine cgas ha

1

Establishing Stable cGAS-HA Cell Line

Check if the same lab product or an alternative is used in the 5 most similar protocols
A plasmid encoding HSV1 UL12 M185 SPA containing a 3X FLAG tag at the C-terminus was described previously24 (link). This construct, or a plasmid encoding murine cGAS-HA (Invivogen), was subcloned into the pMXs-IRES-Puro vector and replication incompetent retroviruses were packaged using plat-E cells according to the manufacturer’s instructions (Cell Biolabs). SV40 large T immortalized cGAS−/− MEFs were exposed to supernatants containing cGAS-HA retroviruses and incubated overnight. Two days post transduction, 3 µg/ml puromycin was added to select a stable population of cells expressing cGAS-HA. Supernatants containing empty or UL12 M185 SPA retroviruses and 4 µg/ml polybrene were incubated with cells 5 × 104 MEFs or 2 × 105 BMDM in 12 well dishes for a period of 8 hours. Viral supernatants were then washed off, fresh media was added to wells, and the cells were incubated for the duration of the experiment until lysis.
West A.P., Khoury-Hanold W., Staron M., Tal M.C., Pineda C.M., Lang S.M., Bestwick M., Duguay B.A., Raimundo N., MacDuff D.A., Kaech S.M., Smiley J.R., Means R.E., Iwasaki A, & Shadel G.S. (2015). Mitochondrial DNA Stress Primes the Antiviral Innate Immune Response. Nature, 520(7548), 553-557.
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2

Establishing Stable cGAS-HA Cell Line

Check if the same lab product or an alternative is used in the 5 most similar protocols
A plasmid encoding HSV1 UL12 M185 SPA containing a 3X FLAG tag at the C-terminus was described previously24 (link). This construct, or a plasmid encoding murine cGAS-HA (Invivogen), was subcloned into the pMXs-IRES-Puro vector and replication incompetent retroviruses were packaged using plat-E cells according to the manufacturer’s instructions (Cell Biolabs). SV40 large T immortalized cGAS−/− MEFs were exposed to supernatants containing cGAS-HA retroviruses and incubated overnight. Two days post transduction, 3 µg/ml puromycin was added to select a stable population of cells expressing cGAS-HA. Supernatants containing empty or UL12 M185 SPA retroviruses and 4 µg/ml polybrene were incubated with cells 5 × 104 MEFs or 2 × 105 BMDM in 12 well dishes for a period of 8 hours. Viral supernatants were then washed off, fresh media was added to wells, and the cells were incubated for the duration of the experiment until lysis.
West A.P., Khoury-Hanold W., Staron M., Tal M.C., Pineda C.M., Lang S.M., Bestwick M., Duguay B.A., Raimundo N., MacDuff D.A., Kaech S.M., Smiley J.R., Means R.E., Iwasaki A, & Shadel G.S. (2015). Mitochondrial DNA Stress Primes the Antiviral Innate Immune Response. Nature, 520(7548), 553-557.
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