Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Von Hippel Lindau/VHL Double Nickase Plasmid (m): sc-423671-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Von Hippel Lindau/VHL Double Nickase Plasmid (m) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- Von Hippel Lindau/VHL Double Nickase Plasmid (m) and Von Hippel Lindau/VHL Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Vhl. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Von Hippel Lindau/VHL Antibody (VHL40): sc-135657
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Von Hippel Lindau/VHL Double Nickase Plasmid (m) | sc-423671-NIC | 20 µg | $410.00 | |||
Von Hippel Lindau/VHL Double Nickase Plasmid (m2) | sc-423671-NIC-2 | 20 µg | $410.00 |
Mouse Vhl encodes the von Hippel–Lindau (VHL) tumor suppressor, the substrate recognition component of a Cullin-2 E3 ubiquitin ligase complex that targets hydroxylated HIF-α for ubiquitination and proteasomal degradation under normoxia. Through regulation of HIF-dependent transcription, VHL controls cellular oxygen sensing, angiogenic and metabolic gene programs, erythropoietic signaling, and redox adaptation. Loss or attenuation of VHL function stabilizes HIF-1α/HIF-2α, reshaping glycolytic flux and extracellular matrix remodeling pathways and altering hypoxia-responsive gene expression. These processes are widely used to model hypoxia signaling, metabolic reprogramming, and microenvironmental responses relevant to tumorigenesis and vascular biology in mouse systems.
Von Hippel Lindau/VHL Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Vhl locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Vhl. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt Vhl function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of Vhl-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.