HIF PHD1 Double Nickase Plasmid (h): sc-402824-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HIF PHD1 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
- HIF PHD1 Double Nickase Plasmid (h) and HIF PHD1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EGLN2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HIF PHD1 Antibody (2G3): sc-293220
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HIF PHD1 Double Nickase Plasmid (h) | sc-402824-NIC | 20 µg | $410.00 | |||
HIF PHD1 Double Nickase Plasmid (h2) | sc-402824-NIC-2 | 20 µg | $410.00 |
Human EGLN2 encodes hypoxia-inducible factor prolyl hydroxylase 1 (HIF PHD1), an Fe(II)/2-oxoglutarate–dependent dioxygenase that hydroxylates HIF-α subunits to promote VHL-mediated ubiquitination and proteasomal turnover under normoxia. By coupling oxygen availability to transcriptional programs, PHD1 helps tune HIF-driven pathways controlling cellular metabolism, angiogenic signaling, erythropoiesis, and adaptive stress responses. EGLN2 activity interfaces with mitochondrial function and redox balance through 2-oxoglutarate metabolism, linking oxygen sensing to broader metabolic regulation. Dysregulated EGLN2/PHD1 signaling has been implicated in contexts of altered hypoxia responses and metabolic remodeling observed across multiple disease-relevant models, supporting mechanistic studies of oxygen-dependent gene regulation.
HIF PHD1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EGLN2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EGLN2. 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 EGLN2 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 EGLN2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.