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cGAS knockout NF-κB-SEAP and IRF-Lucia luciferase reporter monocytes

THP1-Dual KO-cGAS pathway

THP1-Dual^™ KO-cGAS cells were generated from THP1-Dual^™ cells by stable knockout of the cGAS gene. They derive from human THP-1 monocytes, a cell line often used to study DNA sensing pathways as they express all the cytosolic DNA sensors identified so far (with the exception of DAI).

THP1-Dual^™ and THP1-Dual^™ KO-cGAS cells stably express two inducible secreted reporter genes: Lucia luciferase and SEAP (secreted embryonic alkaline phosphatase). The Lucia luciferase reporter gene is under the control of an ISG54 (interferon-stimulated gene) minimal promoter in conjunction with five IFN-stimulated response elements. The SEAP gene is driven by an IFN-β minimal promoter fused to five copies of the NF-kB response element. As a result, they allow the simultaneous study of the IFN regulatory factor (IRF) and the NF-kB pathway by assessing the activity of Lucia luciferase and SEAP, respectively. Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI-Luc™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent, and QUANTI-Blue™ Solution, a SEAP detection reagent.

THP1-Dual™ KO-cGAS and THP1-Dual™ cells can be used to study the role of cGAS by monitoring IRF-induced Lucia luciferase activity. THP1-Dual™ KO-cGAS cells are resistant to blasticidin and Zeocin®.

References:

1. Sun L. et al., 2013. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science 339(6121):786-91.
2. Gao P. et al., 2013. Cyclic [G(2’,5’)pA(3’,5’)p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell. 153(5):1094-107.
3. Ablasser A. et al., 2013. cGAS produces a 2’-5’-linked cyclic dinucleotide second messenger that activates STING. Nature. 498(7454):380-4.

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