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| Selection Marker | Blasticidin, N/A, Puromycin |
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Background
CTLA-4 (CD152) is a key inhibitory immune checkpoint receptor that restrains T cell activation by competing with the costimulatory receptor CD28 for the B7 ligands CD80 and CD86. Alternative splicing of the CTLA4 gene produces a soluble isoform, sCTLA4, that lacks the transmembrane domain and is secreted into the extracellular space. Soluble CTLA-4 can modulate immune responses by binding B7 ligands and is implicated in autoimmune disease and tumor immune regulation. Because sCTLA4 contributes to the balance of immune activation and tolerance, it is studied in immunology and cancer immunotherapy research.
Product Description & Applications
The Human sCTLA4 shRNA Lentivirus delivers a validated short hairpin RNA that achieves at least 70% knockdown of the human soluble CTLA-4 isoform (sCTLA4/CD152). The shRNA is expressed from a U6 promoter in a third-generation, self-inactivating lentiviral backbone, with a co-expressed fluorescent reporter (GFP or RFP, optionally with luciferase) and antibiotic selection marker for stable cell line generation. Knockdown is confirmed using a rapid fluorescence-based assay. Applications include loss-of-function studies of soluble CTLA-4 in immune checkpoint regulation, T cell activation, and tumor immunology. Particles are ultra-purified and concentrated to high titer by PEG precipitation and sucrose gradient centrifugation, transducing difficult-to-transfect cells, including primary and thawed cultures.
About This Product
This validated shRNA lentivirus targeting sCTLA4 (NCBI Accession: NM_001037631.3) delivers a 19–20 bp shRNA from a third-generation, self-inactivating lentiviral backbone. Expression is driven from a U6 Pol III promoter, with a constitutively expressed fluorescent reporter (GFP, GFP/Luc, RFP, RFP/Luc) and antibiotic selection marker (Blasticidin, Puromycin) co-expressed from the same vector. VSV-G pseudotyping enables broad cell tropism, including primary, suspension, and cryopreserved cell types.
Knockdown is validated using a proprietary bicistronic fluorescence assay in which the target mRNA is co-expressed fused to RFP alongside the shRNA-GFP construct. At least 70% reduction in RFP signal in GFP-positive cells confirms on-target activity — a more direct functional readout than transcript-level qPCR. Polyclonal stable lines can be generated by antibiotic selection within 10 days, preserving parental cell heterogeneity compared to single-clone CRISPR approaches.
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.
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