THP1-Lucia™ ISG Cells
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IRF Luciferase Reporter Monocytes
3-7 x 10e6 cells
Interferon Regulatory Factor-Inducible Reporter Monocytes
THP1-Lucia™ ISG cells were derived from the human monocytic cell line THP-1, which represents a model of choice to study the activation and signaling of CDSs.
Indeed, THP-1 cells have been shown to express all the CDSs identified so far [1-3], with the exception of DAI .
THP1-Lucia™ ISG cells express the secreted luciferase (Lucia) reporter gene under the control of an IRF-inducible promoter. This composite promoter is comprised of five IFN-stimulated response elements (ISRE) fused to an ISG54 minimal promoter, which is unresponsive to activators of the NF-kB or AP-1 pathways.
As a result, THP1-Lucia™ ISG cells allow the monitoring of the IRF pathway by determining the activity of Lucia luciferase. The levels of IRF-induced Lucia luciferase in the cell culture supernatant are readily assessed with QUANTI-Luc™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent.
THP1-Lucia™ ISG cells are resistant to Zeocin®.
THP1-Lucia™ ISG cells are highly responsive to STING and CDS ligands, such as transfected double-stranded nucleic acids.
1. Zhang Z. et al., 2011. The helicase DDX41 senses intracellular DNA mediated by the adaptor STING in dendritic cells. Nat Immunol. 12(10):959-65.
2. Veeranki S. et al., 2011. IFI16 protein mediates the anti-inflammatory actions of the type-I interferons through suppression of activation of caspase-1 by inflammasomes. PLoS One. 6(10):e27040.
3. Arakawa R. et al., 2010. Characterization of LRRFIP1. Biochem Cell Biol. 88(6):899-906.
4. Lippmann J. et al., 2010. IFNbeta responses induced by intracellular bacteria or cytosolic DNA in different human cells do not require ZBP1 (DLM-1/DAI). Cell Microbiol. 10(12):2579-88.
Cells were transfected with CDS ligands (1 µg/ml of HSV-60, HSV-60c, VACV-70, VACV-70c, ISD, ISD Control or 0.3 µg/ml pCpGfree-giant, poly(dA:dT), poly(dA), poly(dG:dC) using LyoVec™ or directly stimulated with 10 µg/ml c-di-AMP or c-di-GMP.
After 24h incubation, the levels of IRF-induced Lucia luciferase were determined using QUANTI-Luc™.
Antibiotic resistance: Zeocin®
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)
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- 1 vial containing 3-7 x 106 cells
- 1 ml of Zeocin® (100 mg/ml)
- 1 ml of Normocin™ (50 mg/ml)
- 1 tube of QUANTI-Luc™ 4 Reagent, a Lucia luciferase detection reagent (sufficient to prepare 25 ml)
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