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.
Ferrochelatase Double Nickase Plasmid (h): sc-402043-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ferrochelatase 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
- Ferrochelatase Double Nickase Plasmid (h) and Ferrochelatase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FECH. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ferrochelatase Antibody (A-3): sc-377377
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ferrochelatase Double Nickase Plasmid (h) | sc-402043-NIC | 20 µg | $410.00 | |||
Ferrochelatase Double Nickase Plasmid (h2) | sc-402043-NIC-2 | 20 µg | $410.00 |
Human FECH encodes ferrochelatase, the terminal enzyme of heme biosynthesis that catalyzes insertion of ferrous iron into protoporphyrin IX to form heme in the mitochondrial matrix. This activity supports oxidative metabolism by supplying heme prosthetic groups for hemoglobin, myoglobin, cytochromes, catalases, and nitric oxide synthases. FECH function integrates with mitochondrial iron handling, porphyrin metabolism, and redox homeostasis, making it central to cellular respiration and reactive oxygen species management. Dysregulation of FECH is linked to porphyrin accumulation and heme deficiency phenotypes, including inherited porphyrias such as erythropoietic protoporphyria, and is studied in contexts of anemia, mitochondrial dysfunction, and photosensitivity-associated cellular stress.
Ferrochelatase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FECH locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FECH. 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 FECH 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 FECH-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.