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.
CYP3A5 Double Nickase Plasmid (h): sc-401306-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP3A5 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
- CYP3A5 Double Nickase Plasmid (h) and CYP3A5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CYP3A5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: CYP3A5 Antibody (F18 P3 B6): sc-53616
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP3A5 Double Nickase Plasmid (h) | sc-401306-NIC | 20 µg | $410.00 | |||
CYP3A5 Double Nickase Plasmid (h2) | sc-401306-NIC-2 | 20 µg | $410.00 |
CYP3A5 encodes a cytochrome P450 monooxygenase that catalyzes oxidative metabolism of diverse xenobiotics and endogenous substrates, contributing to phase I biotransformation in the endoplasmic reticulum. As part of hepatic and extrahepatic drug-metabolizing enzyme networks, CYP3A5 influences pathways governing steroid and lipid homeostasis as well as clearance of environmental chemicals. Interindividual variation in CYP3A5 expression and activity is a major determinant of metabolic phenotype and can alter exposure to CYP3A substrates in cellular and organoid models. Dysregulated CYP3A5-dependent metabolism has been investigated in contexts such as toxicant susceptibility and disease-associated changes in hepatic function and inflammation-related transcriptional programs.
CYP3A5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CYP3A5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CYP3A5. 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 CYP3A5 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 CYP3A5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.