RAW-Lucia™ ISG-KO-IFI16 Cells
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Murine RAW 264.7 macrophages - IFI16 Knockout IRF-reporter cells
3-7 x 10e6 cells
IFI16 Knockout IRF-Inducible Lucia luciferase reporter mouse macrophages
RAW-Lucia™ ISG-KO-IFI16 cells were generated from the RAW-Lucia™ ISG cell line, which is derived from the murine RAW 264.7 macrophage cell line, through the stable knockout of murine ortholog of IFI16, p204 (also known as IFI204, PYHIN2).
RAW-Lucia™ ISG-KO-IFI16 cells express a secreted reporter gene, Lucia luciferase, under the control of the I-ISG54 promoter which is comprised of the IFN-inducible ISG54 promoter enhanced by a multimeric ISRE.
RAW 264.7 have been reported to express several CDSs, including IFI16 . RAW-Lucia™ ISG-KO-IFI16 cells can be used to study the importance of IFI16 in the induction of type I IFNs in response to nucleic acids. They allow the monitoring of IRF activation by determining the activity of Lucia luciferase. The levels of IRF-induced Lucia in the cell culture supernatant can be easily monitored using QUANTI-Luc™, a Lucia luciferase detection reagent.
RAW-Lucia™ ISG-KO-IFI16 cells are resistant to Zeocin™.
1. 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.
Stimulation of RAW-Lucia™ ISG-KO-IFI16 and RAW-Lucia™ ISG cells (parental cell line) with poly(dA:dT)/LyoVec™ (1 µg/ml), VACV70/LyoVec™ (1 µg/ml), poly(I:C)/LyoVec™ (1 µg/ml), and 2'3'-cGAMP (3 µg/ml). Mouse IFN-α (1x104 U/ml) and IFN-β (1x104 U/ml) serve as positive controls. Non-induced cells (NI) have been included as a negative control. After a 24h incubation, IRFactivation was determined by measuring the relative light units (RLUs) in a luminometer using QUANTI-Luc™, a Lucia luciferase detection reagent. The IRF induction of each ligand is expressed relative to that of mIFN-β at 1x104 U/ml (taken as 100%).
Antibiotic resistance: Zeocin™
Growth Medium: DMEM, 4.5 g/l glucose, 10% fetal bovine serum (FBS), 100 µg/ml Normocin™, 2 mM L-glutamine
IFI16/p204 knockout is verified by functional assays and DNA sequencing to confirm frameshift mutation/deletion.
The cells are guaranteed mycoplasma-free.
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