| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Reporter | |
| Selection Marker | Blasticidin, GFP (constitutively expressed), Hygromycin, Puromycin, RFP (constitutively expressed), Zeocin |
| Shipping | |
| Species |
Background
CD16A (FcγRIIIa) is a low-affinity Fc receptor for immunoglobulin G expressed on natural killer cells and other effector cells. By engaging the Fc region of antibodies bound to a target cell, CD16A triggers antibody-dependent cellular cytotoxicity (ADCC), a key mechanism by which therapeutic antibodies eliminate tumor and virally infected cells. Receptor engagement raises intracellular calcium and activates the transcription factor NFAT, providing a measurable transcriptional readout of effector-cell activation. A common polymorphism produces high-affinity (V158) and low-affinity (F158) allotypes that influence the strength of ADCC, making CD16A a central focus of antibody-engineering and immuno-oncology research.
Product Description & Applications
The CD16A/NFAT Reporter Lentivirus is an immunotherapy reporter system for quantifying antibody-dependent cellular cytotoxicity. A receptor lentivirus constitutively expresses the human CD16A high-affinity V158 allotype with antibiotic selection, intended for use alongside a companion low-affinity F158 product as a control. A reporter lentivirus carries NFAT response elements driving a dual reporter, secreted Gaussia luciferase and a fluorescent protein. Sequential transduction and selection generates a dual-stable effector cell line, such as Jurkat, that responds to antibody-coated target cells.
When CD16A engages target-bound IgG, intracellular calcium rises and activates NFAT, driving the reporters. Secreted Gaussia luciferase allows kinetic sampling of media without lysis, while fluorescence supports microscopy and flow cytometry. The VSV-G-pseudotyped, PEG- and sucrose-gradient-purified particles transduce difficult-to-transfect cells, supporting ADCC potency testing and antibody screening.
About This Product
This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human ADCC Assay 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, 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.
Can't find the lentiviral construct you need, or want to adjust key design elements? Contact us to discuss custom LV design and optional add-ons.
Common customization requests
- Insert / payload: replace the gene/sequence, swap to a different isoform, add mutations, or optimize cloning features.
- Expression design: change promoter (e.g., CMV/EF1α/PGK), add enhancers, or adjust regulatory elements.
- Reporters: add/swap GFP/RFP/mCherry/luciferase (single or dual reporters where applicable).
- Selection markers: add/swap puromycin/blasticidin/neomycin or fluorescent selection options.
- Vector format: switch between OE, shRNA, CRISPR (sgRNA/Cas systems), or control vectors (where supported).
Add-ons you can request
- Control viruses: empty vector, non-targeting shRNA, reporter-only controls, or matched backbone controls.
- Packaging / format: concentration options, aliquoting, or custom fill volume for screening workflows.
- Documentation: construct map/sequence confirmation package (as available) and batch documentation.
What to include in your request
- Target cell type/model (cell line or primary cells) and intended readout (reporter, knockdown, OE, etc.)
- Insert sequence (FASTA) or reference ID, plus any required tags/mutations
- Promoter, reporter, and selection marker preferences
- Desired scale and preferred format (aliquots / concentration requests)
Email us at support@biohippo.com or use the Talk to a Scientist request form.
