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Murine STING knockin dual reporter monocytes

STING signaling in THP1-Dual™ KI-mSTING cells

THP1-Dual™ KI-mSTING cells stably express the wild-type murine STING (stimulator of interferon genes, mSTING; also known as TMEM173) gene. They were generated by knockin (KI) of the mSTING intronless coding sequence (from ATG to TGA) into THP1-Dual™ KO-STING cells, which derive from the human THP-1 monocytes.

STING is a cytosolic pattern recognition receptor (PRR) that acts as a direct sensor of cyclic dinucleotides (CDNs) [1]. The binding of CDNs to STING leads to TANK-binding-kinase-I (TBK1)-mediated interferon regulatory factor (IRF3) activation and type I interferon (IFN) production [1] and to NF-κB-dependent inflammatory cytokine production [2].

THP1-Dual™ KI-mSTING cells feature two reporter genes allowing the simultaneous study of the IRF pathway, by monitoring the activity of an inducible secreted Lucia luciferase, and the NF-κB pathway by monitoring the activity of an inducible SEAP (secreted embryonic alkaline phosphatase). Lucia luciferase and SEAP activities are readily assessable in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia and QUANTI-Blue™ Solution detection reagents, respectively.

As expected, THP1-Dual™ KI-mSTING cells exhibit a robust response to STING agonists such as 2’3’-cGAMP and 3’3’-cGAMP. In addition, these cells respond to DMXAA (5,6-Dimethylxanthenone 4-acetic acid), a potent activator of murine STING-dependent IFN responses, that does not activate human STING [3]. These cells are resistant to Blasticidin and Zeocin™.

Features of THP1-Dual™ KI-mSTING Cells:

Knockin of the mSTING gene
No functional hSTING expression
IRF activation readily assessable using the Lucia luciferase reporter gene
NF-κB activation readily assessable using the SEAP reporter gene

Validation of THP1-Dual™ KI-mSTING Cells:

Murine STING knockin verified by PCR, DNA sequencing, and Western blot
Functionally tested

References:

1. Sun L. et al., 2013. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science. 339:786-91.
2. Ishikawa H. & Barber G.N. 2008. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455:674-8.
3. Zhang H. et al., 2015. Rat and human STINGs profile similarly towards anticancer/antiviral compounds. Sci Rep. 5:18035.

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