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 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™, 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%).

Specifications

Antibiotic resistance: Zeocin™

Growth Medium: DMEM, 4.5 g/l glucose, 10% fetal bovine serum (FBS), 100 µg/ml Normocin™, 2 mM L-glutamine

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

The cells are guaranteed mycoplasma-free.

This product is covered by a Limited Use License (See Terms and Conditions).

Contents

• 1 vial containing 3-7 x 10⁶ cells
• 100 μl Zeocin™ (100 mg/ml)
• 1 ml Normocin™ (50 mg/ml)
• 1 pouch of QUANTI-Luc™ (Luciferase detection medium)

Shipped on dry ice