RAW-Lucia™ ISG-KO-TRIF Cells

TRIF knockout IRF-inducible Lucia luciferase reporter mouse macrophages

RAW-Lucia^™ ISG-KO-TRIF cells were generated from RAW-Lucia^™ ISG cells through the stable knockout of the TIR-domain-containing adapter-inducing interferon-β (TRIF) gene. These cells derive from the murine RAW 264.7 macrophage cell line, which has been reported to express many pattern recognition receptors (PRRs), including the Toll-like receptors TLR3 and TLR4 [1-3], and their signaling partners. Both TLRs signal through the adapter protein TRIF to induce the activation of interferon regulatory factor (IRF)-3  and the subsequent production of type I interferons.

RAW-Lucia^™ ISG-KO-TRIF and RAW-Lucia^™ ISG cells can be used to study the role of TRIF in a mouse macrophage cell line by monitoring of IRF-induced Lucia luciferase activity. They express the gene for secreted Lucia luciferase under the control of the I-ISG54 promoter which is comprised of the IFN-inducible ISG54 promoter enhanced by a multimeric ISRE. 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-TRIF cells are resistant to Zeocin®.

References:

1. Naiki Y. et al., 2005. Transforming growth factor-beta differentially inhibits MyD88-dependent, but not TRAM- and TRIF-dependent, lipopolysaccharide-induced TLR4 signaling. J Biol Chem. 280(7):5491-5.
2. Goriely S. et al., 2006. Interferon regulatory factor 3 is involved in Toll-like receptor 4 (TLR4)- and TLR3-induced IL-12p35 gene activation. Blood. 107(3):1078-84.
3. Shi CS. &, Kehrl JH., 2008. MyD88 and Trif target Beclin 1 to trigger autophagy in macrophages. J Biol Chem. 283(48):33175-82.

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
TRIF knockout is verified by PCR 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)