RAW-Lucia™ ISG-KO-STING Cells

STING knockout IRF-inducible Lucia luciferase reporter mouse macrophages

RAW-Lucia™ ISG-KO-STING 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 STING gene.
RAW-Lucia™ ISG-KO-STING 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 cGAS [1]. RAW-Lucia™ ISG-KO-STING cells 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-STING cells are resistant to Zeocin®.

Reference:

1. Lam E. et al., 2014. Adenovirus Detection by the cGAS/STING/TBK1 DNA Sensing Cascade. J Virol. 88(2):974-81.

Figures

Analysis of lysates from the RAW-Lucia™ ISG (WT) and RAW-Lucia™ ISG-KO-STING (KO) cells using Anti-STING, followed by an HRP-conjugated anti-rabbit secondary antibody. The arrow indicates the expected band for the murine STING protein (43 kDa).

Response of RAW-Lucia™ ISG-KO-STING cells to various stimuli. RAW-Lucia™ ISG-KO-STING and RAW-Lucia™ ISG cells (parental cell line) were incubated with c-di-AMP (3 µg/ml), 2'3'-cGAMP (3 µg/ml), poly(dA:dT)/LyoVec™ (1 µg/ml), and poly(I:C) HMW/LyoVec™ (1 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:
STING 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 Zeocin® (100 mg/ml)
• 1 ml 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)