Frataxin Double Nickase Plasmid (h): sc-401628-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Frataxin 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
- Frataxin Double Nickase Plasmid (h) and Frataxin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FXN. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Frataxin Antibody (G-12): sc-518079
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Frataxin Double Nickase Plasmid (h) | sc-401628-NIC | 20 µg | $410.00 | |||
Frataxin Double Nickase Plasmid (h2) | sc-401628-NIC-2 | 20 µg | $410.00 |
Human FXN encodes frataxin, a mitochondrial matrix protein required for iron–sulfur (Fe–S) cluster biogenesis and regulation of mitochondrial iron handling. Frataxin supports the ISC assembly machinery to maintain activities of Fe–S–dependent enzymes involved in oxidative phosphorylation, aconitase function, and cellular redox homeostasis. Reduced FXN function is linked to mitochondrial dysfunction, oxidative stress, and altered energy metabolism, making it a key node in pathways governing mitochondrial quality control and iron metabolism. FXN is widely studied in models of neurodegeneration and cardiometabolic stress to connect mitochondrial bioenergetics with iron-driven reactive oxygen species.
Frataxin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FXN locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FXN. 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 FXN 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 FXN-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.