CYP7A1 Double Nickase Plasmid (m): sc-419932-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP7A1 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
- CYP7A1 Double Nickase Plasmid (m) and CYP7A1 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Cyp7a1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CYP7A1 Antibody (E-10): sc-518007
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP7A1 Double Nickase Plasmid (m) | sc-419932-NIC | 20 µg | $410.00 | |||
CYP7A1 Double Nickase Plasmid (m2) | sc-419932-NIC-2 | 20 µg | $410.00 |
Mouse Cyp7a1 encodes cholesterol 7α-hydroxylase (CYP7A1), a liver-enriched cytochrome P450 enzyme that catalyzes the rate-limiting step in the classic bile acid synthesis pathway from cholesterol. By controlling conversion of cholesterol to primary bile acids, CYP7A1 regulates bile acid pool size, hepatic cholesterol homeostasis, and downstream lipid and glucose metabolism through bile acid–responsive signaling networks. Cyp7a1 activity intersects with nuclear receptor pathways including FXR and LXR that coordinate feedback regulation of bile acid biosynthesis and transport. Altered CYP7A1 expression or function is widely used as a mechanistic entry point for studying cholestatic phenotypes, dyslipidemia, and diet-induced metabolic stress in mouse models.
CYP7A1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Cyp7a1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Cyp7a1. 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 Cyp7a1 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 Cyp7a1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.