CYP8B1 Double Nickase Plasmid (h): sc-403383-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP8B1 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
- CYP8B1 Double Nickase Plasmid (h) and CYP8B1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYP8B1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CYP8B1 Antibody (M15-P3B7): sc-101387
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP8B1 Double Nickase Plasmid (h) | sc-403383-NIC | 20 µg | $410.00 | |||
CYP8B1 Double Nickase Plasmid (h2) | sc-403383-NIC-2 | 20 µg | $410.00 |
CYP8B1 encodes sterol 12α-hydroxylase, a microsomal cytochrome P450 enzyme that catalyzes a key hydroxylation step in the classic bile acid biosynthesis pathway, influencing the cholic acid to chenodeoxycholic acid ratio. Through this activity, CYP8B1 helps shape the hepatic bile acid pool, thereby affecting enterohepatic circulation, lipid absorption, and metabolic signaling through bile acid–responsive nuclear receptors and GPCR pathways such as FXR and TGR5. Altered CYP8B1 expression or activity has been associated with dysregulated bile acid composition and broader metabolic phenotypes relevant to liver and cardiometabolic research. CYP8B1 is therefore a useful target for mechanistic studies of hepatic metabolism, bile acid signaling networks, and gene–environment interactions in hepatocytes and relevant model systems.
CYP8B1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYP8B1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYP8B1. 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 CYP8B1 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 CYP8B1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.