The TLR3/NF-κB Luciferase Reporter HEK293 Stable Cell Line is a robust innate immunity and inflammation assay platform designed for quantitative analysis of TLR3-mediated NF-κB signaling. This cell line is derived from human HEK293 cells and is stably engineered to express full-length human Toll-like receptor 3 (TLR3) together with a firefly luciferase reporter gene driven by an NF-κB response element.
This NF-κB luciferase reporter cell line enables sensitive and reproducible detection of TLR3 activation in response to viral RNA mimetics. Upon stimulation with poly(I:C), TLR3 engagement triggers the TRIF-dependent signaling pathway, resulting in NF-κB nuclear translocation and a strong, dose-dependent increase in luciferase activity. The assay performance has been functionally validated using TLR3 ligand assays, making it well suited for innate immune signaling studies and inflammation-focused drug screening.
This stable reporter cell line was generated by co-transfection of a pCMV-TLR3 expression vector, an NF-κB–driven firefly luciferase reporter construct, and an antibiotic resistance vector, followed by antibiotic selection and clone screening. Selected clones exhibit robust poly(I:C)-induced luciferase responses, ensuring high assay sensitivity and reproducibility.
TLR3/NFkB Luciferase Reporter HEK293 Stable Cell Line
Q1: How does poly(I:C) activate NF-κB?
Poly(I:C) is a synthetic analog of viral double-stranded RNA and a well-established TLR3 agonist. Upon endosomal uptake, poly(I:C) binds to TLR3 and initiates TRIF-dependent signaling, leading to phosphorylation events that promote NF-κB nuclear translocation. Activated NF-κB then drives transcription of inflammatory target genes, which can be quantitatively measured using an NF-κB luciferase reporter assay.
