| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Reporter | |
| Selection Marker | Blasticidin, GFP (constitutively expressed), Hygromycin, Puromycin, RFP (constitutively expressed), Zeocin |
| Shipping | |
| Species |
Background
TIM-3 (T cell immunoglobulin and mucin domain-containing protein 3, encoded by HAVCR2) is a co-inhibitory checkpoint receptor expressed on exhausted T cells, regulatory T cells, NK cells, and myeloid cells. Engagement by ligands such as galectin-9, phosphatidylserine, CEACAM1, and HMGB1 dampens T cell receptor signaling and effector function, contributing to immune tolerance and T cell exhaustion in chronic infection and cancer. Because TIM-3 engagement suppresses NFAT-dependent transcription downstream of the T cell receptor, NFAT-driven reporters provide a quantitative readout of TIM-3 signaling, and TIM-3 is an actively pursued checkpoint target in immuno-oncology.
Product Description & Applications
The TIM3/NFAT Reporter Lentivirus is a two-vial immunotherapy reporter system for studying TIM-3 checkpoint signaling. One vial constitutively expresses the human TIM-3 receptor with an antibiotic selection marker, while the other carries tandem NFAT response elements driving a dual reporter combining secreted Gaussia luciferase (GLuc) and a fluorescent protein (GFP or RFP). Sequential transduction and selection generate a dual-stable effector cell line that responds quantitatively to receptor engagement. Secreted GLuc accumulates in conditioned media for kinetic, lysis-free sampling, while the fluorescent reporter enables microscopy and flow cytometry. Applications include evaluating immunotherapies targeting T cell exhaustion and characterizing TIM-3-mediated immune regulation. Supplied as high-titer, VSV-G-pseudotyped third-generation lentiviral particles purified by PEG precipitation and sucrose gradient centrifugation.
About This Product
This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human TIM3 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.
