RAW-Lucia™ ISG-KO-IFI16 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 the 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 [1]. 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™ 4 Lucia/Gaussia, a Lucia and Gaussia luciferase detection reagent.

RAW-Lucia™ ISG-KO-IFI16 cells are resistant to Zeocin®.

Reference:

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.

Figures

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%).

Specifications

Antibiotic resistance: Zeocin®

Growth Medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10% (v/v) heat-inactivated fetal bovine serum, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml Normocin™ supplemented with Zeocin® 

Quality Control: 

IFI16/p204 knockout is verified by functional assays and DNA sequencing to confirm frameshift mutation/deletion.

The cells are guaranteed mycoplasma-free.

Contents

• 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)

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

Published : February 2022