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 (h): sc-400528-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Von Hippel Lindau/VHL Double Nickase Plasmid (h) 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 (h) and Von Hippel Lindau/VHL Double Nickase Plasmid (h2) 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 (h) | sc-400528-NIC | 20 µg | $410.00 | |||
Von Hippel Lindau/VHL Double Nickase Plasmid (h2) | sc-400528-NIC-2 | 20 µg | $410.00 |
VHL encodes the von Hippel–Lindau tumor suppressor, the substrate recognition component of a Cullin 2–RING E3 ubiquitin ligase complex that targets hydroxylated HIF-α for ubiquitination and proteasomal degradation under normoxia. By coupling oxygen sensing to transcriptional control of angiogenesis, metabolism, erythropoiesis, and extracellular matrix programs, VHL helps maintain cellular homeostasis and appropriate hypoxic responses. Loss or dysfunction of VHL disrupts HIF signaling and rewires pathways linked to vascular biology, mitochondrial metabolism, and proteostasis. VHL perturbation is closely associated with inherited VHL disease and is frequently studied in renal cell carcinoma models to interrogate hypoxia-driven tumor biology and microenvironmental adaptation.
Von Hippel Lindau/VHL Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the VHL locus in human 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.