cytochrome b5 Double Nickase Plasmid (m): sc-430951-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- cytochrome b5 Double Nickase Plasmid (m) 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 (m) and cytochrome b5 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Cyb5a. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
cytochrome b5 Double Nickase Plasmid (m) | sc-430951-NIC | 20 µg | $410.00 |
Mouse Cyb5a encodes cytochrome b5, a membrane-associated heme protein that shuttles electrons to cytochrome P450 enzymes, fatty acid desaturases, and elongation systems in the endoplasmic reticulum. Through these interactions, cytochrome b5 supports oxidative metabolism of xenobiotics and endogenous substrates, steroid and lipid biosynthesis, and redox homeostasis. Cyb5a activity can influence cellular responses to oxidative stress and the metabolic handling of drugs and hormones, linking it to pathways relevant for hepatic function and systemic lipid balance. Altered cytochrome b5–dependent electron transfer has been associated with dysregulated lipid metabolism and perturbed P450-mediated oxidation, making Cyb5a a useful node for mechanistic studies in metabolic and toxicological models.
cytochrome b5 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Cyb5a locus in mouse 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.