h/m HIF1a shRNA Lentivirus

Background

HIF1A encodes hypoxia-inducible factor 1-alpha, the oxygen-sensitive subunit of the HIF-1 transcription factor that coordinates the cellular response to low oxygen. Under normal oxygen levels, prolyl hydroxylases modify HIF-1α, marking it for von Hippel-Lindau-mediated degradation. Under hypoxia, this degradation is suppressed, allowing HIF-1α to dimerize with HIF-1β and activate genes that promote angiogenesis, glycolysis, erythropoiesis, and metabolic adaptation. HIF-1 signaling is essential for development and ischemic tissue responses, and it is frequently hyperactivated in solid tumors, where it drives angiogenesis, metabolic reprogramming, and resistance to therapy. HIF-1α is therefore a key focus of cancer and cell biology research.

Product Description & Applications

The h/m HIF1a shRNA Lentivirus delivers validated short hairpin RNA targeting human and mouse HIF1A for stable gene knockdown. Each shRNA is validated to achieve at least 70% reduction of HIF1A using a fluorescence-based assay that provides a direct functional readout rather than transcript-level qPCR. The particles are ultra-purified and concentrated to high titer by PEG precipitation and sucrose gradient centrifugation, and efficiently transduce difficult-to-transfect cells, including primary and thawed cultures. A co-expressed fluorescent reporter (GFP or RFP, optionally with luciferase) and antibiotic selection marker (puromycin or blasticidin) enable stable line generation. A shRNA set option supplies a mix of two independent validated shRNAs plus matched scrambled-control lentivirus for loss-of-function studies of hypoxia signaling and tumor biology.

About This Product

This validated shRNA lentivirus targeting HIF1A (NCBI Accession: NM_001530) 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 (Puromycin, Blasticidin) 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.