CLEC4E/NFκB Reporter Lentivirus

Background

CLEC4E, also known as Mincle, is a C-type lectin receptor expressed mainly on myeloid cells such as macrophages and dendritic cells. It functions as a pattern-recognition receptor that senses pathogen-associated molecular patterns, including mycobacterial glycolipids, as well as damage-associated ligands released from dying cells. Ligand binding triggers signaling through the FcRgamma adaptor and the Syk-CARD9 axis, leading to activation of the NF-κB family of transcription factors and the production of pro-inflammatory cytokines. Through this pathway, CLEC4E shapes innate immune responses to infection and tissue damage, making it an important focus in research on antifungal and antimycobacterial immunity and inflammation.

Product Description & Applications

The CLEC4E/NF-κB Reporter Lentivirus is an all-in-one immunotherapy reporter system for studying CLEC4E (Mincle) signaling in immune cells. The construct constitutively expresses the human CLEC4E receptor and couples ligand recognition to NF-κB-dependent transcription of dual reporters: secreted Gaussia luciferase and a fluorescent protein (GFP or RFP), enabling real-time monitoring by luminometry and fluorescence.

Transducing target cells with the high-titer lentivirus establishes a stable, pathway-specific reporter cell line. Applications include studying innate immunity, characterizing pathogen-sensing mechanisms, and evaluating CLEC4E-targeted compounds. Supplied as third-generation, VSV-G-pseudotyped particles purified by PEG precipitation and sucrose gradient centrifugation, the product is effective in difficult-to-transfect primary and thawed cells.

About This Product

This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human CLEC4E under a constitutive promoter with antibiotic selection, and a Vial 2 Reporter Lentivirus encoding tandem NFAT (or NF-κB) response elements driving a dual reporter (GFP, GFP-P2A-GLuc, GLuc, GLuc-P2A-GFP, GLuc-P2A-RFP, RFP, RFP-P2A-GLuc). Sequential transduction and selection generates a dual-stable effector cell line that responds quantitatively to receptor stimulation with a ratiometric fluorescent + bioluminescent readout.

Secreted Gaussia luciferase (where included) accumulates in conditioned media, enabling kinetic sampling without cell lysis. The combined fluorescent and luminescent outputs allow parallel microscopy-based visualization and plate-reader luminometry from the same cell population — providing assay redundancy and flexibility for potency testing formats compliant with regulatory expectations for cell-based functional assays.