HEK-Lucia™ RIG-I Cells

Human RIG-I lucia luciferase reporter HEK293 cells

HEK-Lucia™ RIG-I cells were generated from HEK-Lucia™ Null cells, HEK293-derived cells that stably express the secreted Lucia luciferase reporter gene.

This reporter gene is under the control of an IFN‑inducible ISG54 promoter enhanced by a multimeric IFN‑stimulated response elements (ISRE).

HEK-Lucia™ RIG-I cells stably express high levels of human RIG-I (also known as Ddx58), and respond strongly to cytosolic double-stranded RNAs with an uncapped 5'-di/triphosphate end [1,2] such as 3p-hpRNA and 5’ppp‑dsRNA (see DATA sheet).

HEK-Lucia™ RIG-I and HEK-Lucia™ Null cells can be used to study the role of RIG-I by monitoring IRF-induced Lucia luciferase activity.

The levels of IRF-induced Lucia in cell culture supernatants can be easily monitored using QUANTI-Luc™, a Lucia luciferase detection reagent.

HEK-Lucia™ RIG-I cells are resistant to blasticidin, G418, hygromycin, and  Zeocin™.
They should be maintained in growth medium supplemented with blasticidin and Zeocin™.

Note: HEK-Lucia™ RIG-I and HEK-Lucia™ Null cells endogenously express NOD1, TLR3 and TLR5.

References

1. Gebhardt A. et al., 2017. Discrimination of Self and Non-Self Ribonucleic Acids. Journal of Interferon & Cytokine Research 37: 184-97.
2. Pichlmair A. et al., 2006. RIG-I-mediated antiviral responses to single-stranded RNA bearing 5’-phosphates. Science 314:997-1001.

Figures

HEK-Lucia™ Null and HEK-Lucia™ RIG-I cells were stimulated with 1 μg/ml 5’ppp-dsRNA or 3p-hpRNA (pannel 1), or with 100 ng/ml of Poly(I:C) HMW/Lyovec™, 100 ng/ml of Poly(I:C) LMW/Lyovec™, or 100 ng/ml of Poly(dA:dT)/Lyovec™ (pannel 2). After overnight incubation, the ISG response was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™ and expressed as relative light units (RLUs).

Specifications

Antibiotic resistance: Blasticidin, G418, hygromycin, and  Zeocin™

Growth medium: DMEM, 4.5 g/l glucose, 2 mM L-glutamine, 10%heat-inactivated fetal bovine serum, 100 μg/ml Normocin™ , Pen-Strep (100 U/ml-100 µg/ml)

Quality Control:
- The expression of the human RIG-I gene has been confirmed by qRT-PCR.
- The reporter activity has been validated by stimulating these cells with various RIG-I agonists.
- The cell line stability for 20 passages following thawing has been verified.
- HEK-Lucia™ RIG-I cells are guaranteed mycoplasma-free.

Contents

• 1 vial of HEK-Lucia™ RIG-I cells (3-7 x 10⁶ cells)
• 1 ml of Zeocin™ (100 mg/ml)
• 1 ml of blasticidin (10 mg/ml)
• 1 ml of Normocin™ (50 mg/ml)
• 1 pouch of QUANTI-Luc™ (Lucia luciferase detection medium)

Shipped on dry ice (Europe, USA & Canada)