| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Reporter | |
| Selection Marker | Blasticidin, GFP (constitutively expressed), Hygromycin, Puromycin, RFP (constitutively expressed), Zeocin |
| Shipping | |
| Species |
Background
CTLA-4 (CD152) is a co-inhibitory receptor and a central regulator of immune checkpoint control. Expressed on activated T cells and constitutively on regulatory T cells, it competes with the co-stimulatory receptor CD28 for the shared B7 ligands CD80 and CD86 on antigen-presenting cells, and binds them with higher affinity. By outcompeting CD28 and removing B7 ligands from the cell surface, CTLA-4 dampens T cell activation, reduces calcium-driven NFAT signaling, and enforces peripheral tolerance. Because CTLA-4 restrains anti-tumor immunity, blocking antibodies are a foundational cancer immunotherapy, and NFAT reporters provide a quantitative readout of CTLA-4-mediated suppression and its reversal.
Product Description & Applications
The CTLA4/NFAT Reporter Lentivirus is an all-in-one immunotherapy reporter system for studying CTLA-4-mediated checkpoint signaling in T cells. The construct constitutively expresses the human CTLA-4 receptor; engagement of B7 ligands suppresses T cell activity, lowering cytosolic calcium, reducing NFAT activation, and decreasing dual reporter output of secreted Gaussia luciferase and a fluorescent protein (GFP or RFP). Anti-CTLA-4 antibodies that block receptor engagement restore reporter activity.
Transducing target cells with the high-titer lentivirus establishes a stable, pathway-specific reporter cell line. Applications include studying immune checkpoint suppression and testing checkpoint-blockade agents. Supplied as third-generation, VSV-G-pseudotyped particles purified by PEG precipitation and sucrose gradient centrifugation, effective in primary and thawed cells.
About This Product
This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human CTLA4 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.
