DD4 Double Nickase Plasmid (h): sc-403692-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- DD4 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
- DD4 Double Nickase Plasmid (h) and DD4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AKR1C4. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: DD4 Antibody (WW3): sc-100526
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
DD4 Double Nickase Plasmid (h) | sc-403692-NIC | 20 µg | $410.00 | |||
DD4 Double Nickase Plasmid (h2) | sc-403692-NIC-2 | 20 µg | $410.00 |
AKR1C4 encodes an aldo-keto reductase (AKR1C4; DD4) that catalyzes NADPH-dependent reduction of aldehydes and ketosteroids, contributing to hepatic steroid hormone and bile acid metabolism. DD4 participates in phase I detoxification by modulating the interconversion of active and inactive steroid metabolites and processing reactive carbonyl compounds generated during oxidative stress. Through its roles in carbonyl reduction and steroid homeostasis, AKR1C4 influences metabolic signaling networks linked to lipid handling and hepatic xenobiotic clearance. Altered AKR1C4 expression or activity has been associated with dysregulated steroid metabolite profiles and liver-associated metabolic phenotypes, making it relevant for mechanistic studies of endocrine-metabolic crosstalk.
DD4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AKR1C4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AKR1C4. 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 AKR1C4 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 AKR1C4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.