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| Selection Marker | Blasticidin, N/A, Puromycin |
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Background
Hypoxia-inducible factor 2 alpha (HIF-2α), encoded by EPAS1, is an oxygen-sensitive transcription factor that drives cellular adaptation to low oxygen. Under normoxia it is hydroxylated and degraded; under hypoxia it is stabilized, dimerizes with HIF-1β, and binds hypoxia-response elements to activate target genes. HIF-2α regulates erythropoiesis, angiogenesis, iron metabolism, and stem cell maintenance, with a gene set partly distinct from HIF-1α. Aberrant HIF-2α activity contributes to clear cell renal carcinoma and other tumors, where it promotes growth and vascularization, making EPAS1 an important target in cancer and hypoxia research.
Product Description & Applications
The h/m HIF2a shRNA Lentivirus delivers a validated short hairpin RNA targeting human and mouse EPAS1 (HIF2a) for stable RNA interference. 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 optional blasticidin or puromycin selection. Particles are ultra-purified and concentrated by PEG precipitation and sucrose gradient centrifugation and efficiently transduce difficult-to-transfect cells, including primary and thawed cells. The shRNA is validated for at least 70% knockdown using a fluorescence-based assay.
A shRNA set option supplies a mix of two validated shRNAs plus a scrambled control for loss-of-function studies of hypoxia signaling and tumor biology.
About This Product
This validated shRNA lentivirus targeting EPAS1 (NCBI Accession: NM_001430) 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.
