| Field | Specification |
|---|---|
| Mfr No | |
| Accession Number | |
| Product Type | |
| Promoter | |
| Reporter | |
| Selection Marker | Blasticidin, N/A, Puromycin |
| Shipping | |
| Species |
Background
HIF1A encodes hypoxia-inducible factor 1-alpha, the oxygen-sensitive subunit of the HIF-1 transcription factor that orchestrates the cellular response to low oxygen. Under normoxia, prolyl hydroxylases modify conserved proline residues in HIF-1α, marking it for von Hippel-Lindau-mediated ubiquitination and proteasomal degradation. Under hypoxia, this degradation is blocked, allowing HIF-1α to dimerize with HIF-1β and activate genes governing angiogenesis, glycolysis, erythropoiesis, and metabolic adaptation. HIF-1 signaling is central to development, ischemic disease, and tumor biology, where it drives angiogenesis and metabolic reprogramming. Mutating the regulatory proline and asparagine residues (P402A/P577A/N813A) yields a stabilized protein that remains active under normoxic conditions.
Product Description & Applications
The HIF1A Wild Type & Mutant ORF cDNA Lentivirus delivers the human or mouse HIF1A open reading frame for stable overexpression in mammalian cells, available in wild-type form and as a normoxia-stable mutant (P402A/P577A/N813A) resistant to oxygen-mediated degradation. The ORF is fused to a C-terminal V5 tag and linked via self-cleaving peptides to a fluorescent reporter (GFP or RFP) or luciferase and a drug-selection marker (puromycin or blasticidin); expression is driven by a CMV or EF1a promoter, with PGK driving reporter and selection cassettes in some constructs. The particles efficiently transduce difficult-to-transfect cells, including primary and thawed cultures, supporting stable cell line generation for studies of hypoxia signaling, angiogenesis, and tumor metabolism. ORF integrity is verified by sequencing and expression confirmed by transient transfection.
About This Product
This ORF cDNA lentivirus enables stable overexpression of HIF1A (NCBI Accession: NM_001530, NM_010431) in mammalian cells via a third-generation, VSV-G pseudotyped delivery system. The ORF cDNA is fused to a C-terminal epitope tag (V5, Myc, or HA) and expressed under a strong constitutive promoter (CMV). Reporter and selection marker components (GFP, RFP; Blasticidin, Puromycin) are co-expressed via self-cleaving P2A peptides, enabling independent protein production without fusion-tag artifacts.
Ultra-purification by PEG precipitation and sucrose gradient centrifugation yields high-titer particles suitable for primary cells, suspension cultures, and stem cells. Stable polyclonal cell lines are established within 10–14 days by antibiotic selection or FACS sorting. For in vivo applications, the serum-free formulation and VSV-G envelope support direct administration or further concentration for stereotactic injection.
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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