CYP27A1 Double Nickase Plasmid (h): sc-402836-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP27A1 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
- CYP27A1 Double Nickase Plasmid (h) and CYP27A1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYP27A1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CYP27A1 Antibody (G-2): sc-390974
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP27A1 Double Nickase Plasmid (h) | sc-402836-NIC | 20 µg | $410.00 | |||
CYP27A1 Double Nickase Plasmid (h2) | sc-402836-NIC-2 | 20 µg | $410.00 |
Human CYP27A1 encodes sterol 27-hydroxylase, a mitochondrial cytochrome P450 enzyme that catalyzes oxidation of cholesterol and sterol intermediates during bile acid biosynthesis. By generating 27-hydroxylated sterols, CYP27A1 supports mitochondrial sterol catabolism, contributes to lipid homeostasis, and interfaces with hepatic bile acid, cholesterol transport, and oxysterol signaling pathways. Altered CYP27A1 activity affects sterol and bile acid composition and has been linked to inborn errors of bile acid metabolism such as cerebrotendinous xanthomatosis, with downstream effects on cholesterol deposition and tissue sterol handling. CYP27A1 is therefore widely studied in metabolic biology, mitochondrial P450 function, and oxysterol-mediated regulation of nuclear receptor networks.
CYP27A1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYP27A1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYP27A1. 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 CYP27A1 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 CYP27A1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.