| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Reporter | |
| Selection Marker | Puromycin + Blasticidin |
| Shipping | |
| Species |
Background
SIRPα (SIRPA) is an inhibitory transmembrane receptor expressed predominantly on macrophages and dendritic cells. Its ligand, CD47, is a widely expressed immunoglobulin-superfamily protein that is frequently overexpressed by tumor cells. Engagement of CD47 by SIRPα transmits a "don't eat me" signal that suppresses phagocytosis, allowing cancer cells to evade macrophage-mediated clearance. Blocking the CD47-SIRPα axis with antibodies or SIRPα-Fc fusion proteins restores phagocytic activity, making this interaction a major target in cancer immunotherapy. NF-κB is a key transcription factor activated downstream of innate immune receptors, providing a quantitative readout of pathway engagement in this reporter system.
Product Description & Applications
The SIRPA/NFκB Reporter Lentivirus Set is a two-component immunotherapy reporter system for studying the CD47-SIRPα phagocytosis checkpoint. One lentivirus constitutively expresses the SIRPA receptor; the second carries NF-κB response elements driving a dual reporter of secreted Gaussia luciferase and a fluorescent protein (eGFP or mCherry). Upon receptor engagement, NF-κB activation produces a quantitative luminescent and fluorescent signal. Sequential transduction and selection generate a stable reporter cell line for measuring the potency of SIRPα-blocking antibodies or SIRPα-Fc in "don't eat me" phagocytosis-inhibition assays. Secreted Gaussia luciferase enables non-destructive kinetic sampling from conditioned media. Supplied as high-titer, VSV-G-pseudotyped third-generation particles purified by PEG precipitation and sucrose gradient centrifugation, suitable for primary and thawed cells.
About This Product
This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human Don't Eat Me' 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 (Gaussia Luciferase + eGFP, Gaussia Luciferase + mCherry). 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.
