RAW-Lucia™ ISG Cells
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Mouse Macrophages - CDS Reporter Cells
3-7 x 10e6 cells
Mouse Macrophages - CDS Reporter Cells
RAW-Lucia™ ISG cells were generated from the murine RAW 264.7 macrophage cell line by stable integration of an interferon regulatory factor (IRF)-inducible Lucia luciferase reporter construct.
RAW 264.7 have been reported to express many pattern recognition receptors (PRRs), including the cytosolic DNA sensors (CDS), IFI16 , DDX41, AIM2  and cGAS , the dsRNA sensor RIG-I , and the cyclic dinucleotide sensor STING . Activation of most of these receptors induces the production of type I IFNs through the TBK1-IRF3 axis.
Cell line description:
RAW-Lucia™ ISG cells express the Lucia luciferase gene under the control of an ISG54 minimal promoter in conjunction with five IFN-stimulated response elements. Thus, RAW-Lucia™ ISG cells allow the monitoring of IRF activation by determining the activity of Lucia luciferase. The levels of IRF-induced Lucia luciferase in the cell culture supernatant can be easily monitored using QUANTI-Luc™, a luciferase detection reagent. RAW-Lucia™ ISG cells are resistant to Zeocin™.
RAW-Lucia™ ISG cells are responsive to murine IFN-α and IFN-β but do not respond to their human counterparts. They are also responsive to PRR ligands that trigger the IFN signaling pathway, including CDS ligands, such as transfected double-stranded DNA, RIG-I ligands, such as transfected double-stranded RNA, and STING ligands, such as cGAMP.
Features of RAW-Lucia™ ISG cells:
- Fully functional STING/TBK1/IRF3 signaling pathway
- Readily assessable Lucia luciferase reporter activity
- Functionally tested and guaranteed mycoplasma-free
Applications of RAW-Lucia™ ISG cells:
- Studying the STING/TBK1/IRF3 signaling pathway
- Studying the RIG-I/TBK1/IRF3 signaling pathway
1. Kawasaki T. et al., 2011. Recognition of nucleic acids by pattern-recognition receptors and its relevance in autoimmunity. Immunol Rev. 243(1):61-73.
2. Stein SC & Falck-Pedersen E., 2012. Sensing adenovirus infection: activation of interferon regulatory factor 3 in RAW 264.7 cells. J Virol. 86(8):4527-37.
3. Lam E. et al., 2014. Adenovirus Detection by the cGAS/STING/TBK1 DNA Sensing Cascade. J Virol. 88(2):974-81.
4. Melchjorsen J. et al., 2005. Activation of innate defense against a paramyxovirus is mediated by RIG-I and TLR7 and TLR8 in a cell-type-specific manner. J Virol. 79(20):12944-51.
5. Tanaka Y. & Chen ZJ., 2012. STING specifies IRF3 phosphorylation by TBK1 in the cytosolic DNA signaling pathway. Sci Signal. 5(214):ra20.
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™
These products are covered by a Limited Use License (See Terms and Conditions).Back to the top
- 1 vial containing 5-7 x 106 cells
- 100 μl Zeocin™ (100 mg/ml)
- 1 ml Normocin™ (50 mg/ml)
- 1 pouch of QUANTI-Luc™
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STING (stimulator of interferon genes; also known as TMEM173, MITA, MPYS and ERIS) is essential for the interferon (IFN) response to cytosolic nucleic acids, such as microbial or self-DNA [1, 2], and acts as a direct sensor of cyclic dinucleotides (CDNs) . CDNs are important second messenger molecules in bacteria, affecting numerous responses of the prokaryotic cell. In mammalian cells, CDNs act as agonists of the innate immune response . CDNs bind directly to and activate STING leading to TANK Binding Kinase 1 (TBK1)-dependent interferon regulatory factor 3 (IRF3) activation and type I IFN production. IFNs then activate the JAK-STAT pathway with subsequent activation of IFN-stimulated response elements (ISRE) in the promoters of IFN-stimulated genes (ISG).
1. Wan D. et al., 2020. Research Advances in How the cGAS-STING Pathway Controls the Cellular Inflammatory Response. Front Immunol. 11:615.
2. Ishikawa H. & Barber, G., 2008. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455, 674–678.
3. Burdette D.L. et al., 2011. STING is a direct innate immune sensor of cyclic di-GMP. Nature 478(7370):515-8.
4. Wu J. et al., 2013. Cyclic GMP-AMP is an endogenous second messenger in innate immune signaling by cytosolic DNA. Science 339(6121):826-30.