HFE Double Nickase Plasmid (h): sc-403695-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- HFE 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
- HFE Double Nickase Plasmid (h) and HFE Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting HFE. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: HFE Antibody (C-12): sc-514405
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
HFE Double Nickase Plasmid (h) | sc-403695-NIC | 20 µg | $410.00 | |||
HFE Double Nickase Plasmid (h2) | sc-403695-NIC-2 | 20 µg | $410.00 |
HFE encodes a non-classical MHC class I–like protein that associates with β2-microglobulin and modulates transferrin receptor binding to regulate cellular iron uptake. Through this interaction, HFE helps coordinate iron sensing and systemic iron homeostasis across hepatocytes, enterocytes, and macrophages, influencing hepcidin regulation and iron storage pathways. Disrupted HFE function alters iron trafficking and oxidative stress responses, linking the gene to iron overload phenotypes such as hereditary hemochromatosis and downstream tissue injury mechanisms. HFE is therefore widely used as a model for studying iron metabolism, metal-responsive signaling, and iron-driven inflammatory or fibrotic processes.
HFE Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the HFE locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within HFE. 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 HFE 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 HFE-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.