THP-1-Dual KO-STING cell line穩定敲除STING的THP-1細胞株

STING knockout dual reporter monocytes
THP1-Dual? KO-STING cells were generated from THP1-Dual? cells by stable knockout of the STING gene. STING (stimulator of interferon genes), alternatively known as MPYS, TMEM173, MITA and ERIS is a direct sensor of cyclic dinucleotides (CDNs) [1-3]. These cell lines were derived from human THP_x001e_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-STING cells stably express two inducible secreted reporter genes: IFN-inducible Lucia luciferase and NF_x001e_κB-inducible SEAP (secreted embryonic alkaline phosphatase). They can be used to study the role of STING by monitoring IRF-induced Lucia luciferase activity, using QUANTI_x001e_Luc?, a Lucia? luciferase detection reagent.

THP1-Dual? KO-STING cells are resistant to the selectable markers blasticidin and Zeocin?.



References:

1. Burdette DL. et al., 2011. STING is a direct innate immune sensor of cyclic di-GMP. Nature. 478(7370):515-8.
2. Zhang X. et al., 2013. Cyclic GMP-AMP containing mixed phosphodiester linkages is an endogenous high-affinity ligand for STING. Mol Cell. 51(2):226-35.
3. Barber GN. et al., 2015. STING: infection, inflammation and cancer. Nat Rev Immunol. 15(12):760-70.

Specifications
Antibiotic resistance: Zeocin?, blasticidin

Growth medium:  RPMI 1640, 2 mM L-glutamine, 25 mM HEPES, 10% heat-inactivated fetal bovine serum, 100 μg/ml Normocin?, Pen-Strep (100 U/ml-100 μg/ml)

Quality control:

Biallelic STING knockout is verified by functional assays, PCR and DNA sequencing.

These cells are guaranteed mycoplasma-free.

Description
THP1-Dual? and THP1-Dual? KO-STING 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_x001e_κB response element. As a result, they allow the simultaneous study of the IFN regulatory factor (IRF) pathway, by assessing the activity of Lucia luciferase and the NF-κB pathway, by monitoring the activity of SEAP. Both reporter proteins are readily measurable in the cell culture supernatant when using QUANTI_x001e_Luc?, a Lucia? luciferase detection reagent and QUANTI-Blue?, a SEAP detection reagent.

THP1-Dual? KO-STING and THP1-Dual? cells can be used to study the role of STING by monitoring IRF-induced Lucia luciferase activity. Unlike the parental cells, THP1-Dual? KO_x001e_STING cells exhibit no detectable response to cytosolic DNA and CDNs while retaining the ability to respond to type I IFNs. As expected these cells remain responsive to RIG-I ligands such as transfected Poly(I:C).

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