| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Reporter | |
| Selection Marker | Blasticidin, GFP (constitutively expressed), Hygromycin, Puromycin, RFP (constitutively expressed), Zeocin |
| Shipping | |
| Species |
Background
VISTA (V-domain Ig suppressor of T cell activation) is an immune checkpoint receptor and ligand of the immunoglobulin superfamily, expressed predominantly on myeloid cells and, to a lesser extent, on T cells. VISTA delivers inhibitory signals that suppress T cell activation and proliferation and help maintain peripheral tolerance and a suppressive myeloid compartment. In tumors, VISTA contributes to an immunosuppressive microenvironment and immune evasion, making it an attractive checkpoint target. Because VISTA-mediated inhibition dampens NFAT-dependent transcription downstream of the T cell receptor, NFAT-driven reporters provide a quantitative readout of VISTA checkpoint signaling for immuno-oncology research.
Product Description & Applications
The VISTA/NFAT Reporter Lentivirus is an immunotherapy reporter system for studying VISTA checkpoint signaling. A receptor lentivirus constitutively expresses human VISTA with antibiotic selection, while a reporter lentivirus carries tandem NFAT response elements driving a dual reporter, secreted Gaussia luciferase and a fluorescent protein. Sequential transduction and selection generates a dual-stable cell line whose NFAT-driven reporter output reflects VISTA-modulated signaling.
Secreted Gaussia luciferase accumulates in conditioned media, enabling kinetic sampling without cell lysis, while fluorescence supports microscopy and flow cytometry. The VSV-G-pseudotyped particles are purified by PEG precipitation and sucrose gradient centrifugation and transduce difficult-to-transfect cells, including primary and thawed cultures. The system supports investigation of immune evasion, checkpoint inhibitor testing, and the role of VISTA in myeloid cell suppression.
About This Product
This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human VISTA 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.
