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.
HIF PHD2 Double Nickase Plasmid (h): sc-403334-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HIF PHD2 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 PHD2 Double Nickase Plasmid (h) and HIF PHD2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EGLN1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HIF PHD2 Antibody (H-8): sc-271835
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HIF PHD2 Double Nickase Plasmid (h) | sc-403334-NIC | 20 µg | $410.00 | |||
HIF PHD2 Double Nickase Plasmid (h2) | sc-403334-NIC-2 | 20 µg | $410.00 |
EGLN1 encodes HIF prolyl hydroxylase 2 (PHD2), an oxygen-sensing dioxygenase that hydroxylates HIF-α subunits in an iron- and 2-oxoglutarate–dependent reaction. Under normoxia, this modification promotes VHL-mediated ubiquitination and proteasomal degradation of HIF-α, thereby limiting transcriptional programs that control angiogenesis, glycolytic metabolism, erythropoiesis, and cell survival. PHD2 integrates inputs from oxygen availability, mitochondrial metabolism, and reactive oxygen species to tune hypoxia-inducible signaling. Dysregulated EGLN1/HIF pathway activity has been linked to disorders of oxygen homeostasis and contributes to hypoxia-driven phenotypes relevant to cancer biology, ischemia models, and inflammatory microenvironments.
HIF PHD2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EGLN1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EGLN1. 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 EGLN1 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 EGLN1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.