| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Reporter | |
| Selection Marker | Blasticidin, GFP (constitutively expressed), Hygromycin, Puromycin, RFP (constitutively expressed), Zeocin |
| Shipping | |
| Species |
Background
ICOS (inducible T cell co-stimulator) is a CD28-family co-stimulatory receptor induced on activated T cells that, upon engaging its ligand ICOS-L, enhances T cell activation, cytokine production, and helper T cell function. CD3-zeta is the principal signal-transducing subunit of the T cell receptor complex, carrying ITAM motifs that initiate activation upon antigen recognition. Fusing the ICOS ectodomain to the CD3-zeta signaling chain creates a chimeric receptor that converts ICOS-ligand engagement into a T cell receptor-like activation signal. Both ICOS co-stimulation and CD3-zeta signaling converge on NFAT, a transcription factor that drives effector gene expression, making NFAT reporters a quantitative measure of this engineered signaling.
Product Description & Applications
The ICOS-CD3ζ/NFAT Reporter Lentivirus is a two-component immunotherapy reporter system that models co-stimulatory signaling in T cells. A receptor construct constitutively expresses the human ICOS-CD3ζ fusion protein with an antibiotic selection marker, while a reporter construct drives a dual reporter through tandem NFAT response elements. Receptor engagement activates NFAT, producing a fluorescent (GFP or RFP) and secreted Gaussia luciferase readout.
Sequential transduction and selection generate a stable, pathway-specific effector cell line. Applications include studying T cell activation, immune modulation, and co-stimulatory pathways in immunotherapy. Supplied as high-titer, VSV-G-pseudotyped lentiviral particles, the product is suitable for establishing reporter lines in primary or thawed T cells.
About This Product
This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human ICOS-CD3ζ fusion 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.
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.
