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.
EPAS-1/HIF-2 alpha Double Nickase Plasmid (h): sc-400647-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EPAS-1/HIF-2 alpha 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
- EPAS-1/HIF-2 alpha Double Nickase Plasmid (h) and EPAS-1/HIF-2 alpha Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPAS1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EPAS-1/HIF-2 alpha Antibody (190b): sc-13596
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EPAS-1/HIF-2 alpha Double Nickase Plasmid (h) | sc-400647-NIC | 20 µg | $410.00 | |||
EPAS-1/HIF-2 alpha Double Nickase Plasmid (h2) | sc-400647-NIC-2 | 20 µg | $410.00 |
EPAS1 encodes EPAS-1/HIF-2 alpha, an oxygen-sensitive bHLH-PAS transcription factor that heterodimerizes with ARNT (HIF-1β) to coordinate hypoxia-adaptive gene expression. Under low oxygen, EPAS-1/HIF-2 alpha escapes prolyl hydroxylase–VHL–mediated degradation, accumulates in the nucleus, and activates programs controlling angiogenesis, erythropoiesis, iron metabolism, and metabolic rewiring. EPAS1 signaling intersects with PI3K–AKT–mTOR and MAPK pathways and contributes to cell fate decisions in endothelial and other oxygen-responsive tissues. Dysregulated EPAS1 activity and altered hypoxia signaling are implicated in tumor biology and vascular remodeling, and EPAS1 variants have been associated with oxygen-sensing disorders.
EPAS-1/HIF-2 alpha Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPAS1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPAS1. 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 EPAS1 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 EPAS1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.