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.
cytochrome b5 Double Nickase Plasmid (h): sc-405811-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- cytochrome b5 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
- cytochrome b5 Double Nickase Plasmid (h) and cytochrome b5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYB5A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: cytochrome b5 Antibody (36): sc-130311
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
cytochrome b5 Double Nickase Plasmid (h) | sc-405811-NIC | 20 µg | $410.00 | |||
cytochrome b5 Double Nickase Plasmid (h2) | sc-405811-NIC-2 | 20 µg | $410.00 |
Human CYB5A encodes cytochrome b5, a membrane-anchored hemoprotein that shuttles electrons from NADH–cytochrome b5 reductase to multiple oxidative enzymes in the endoplasmic reticulum and outer mitochondrial membrane. It supports key lipid metabolic pathways, including fatty acid desaturation and elongation, cholesterol and steroid biosynthesis, and xenobiotic oxidation through modulation of cytochrome P450 catalytic cycles. By influencing redox balance and metabolic flux, CYB5A contributes to cellular homeostasis in hepatocytes and steroidogenic tissues where oxidative metabolism is high. Perturbation of CYB5A-dependent electron transfer has been linked to altered drug metabolism and steroidogenic defects, making it a useful node for studying metabolic dysregulation and redox-sensitive signaling.
cytochrome b5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYB5A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYB5A. 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 CYB5A 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 CYB5A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.