Aldose Reductase Double Nickase Plasmid (h): sc-401437-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Aldose Reductase 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
- Aldose Reductase Double Nickase Plasmid (h) and Aldose Reductase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting AKR1B1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Aldose Reductase Antibody (H-6): sc-166918
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Aldose Reductase Double Nickase Plasmid (h) | sc-401437-NIC | 20 µg | $410.00 | |||
Aldose Reductase Double Nickase Plasmid (h2) | sc-401437-NIC-2 | 20 µg | $410.00 |
AKR1B1 encodes human aldose reductase, an NADPH-dependent aldo-keto reductase that catalyzes the reduction of glucose to sorbitol as the first step of the polyol pathway. By consuming NADPH and promoting sorbitol accumulation, aldose reductase can influence cellular redox balance, osmotic stress responses, and downstream fructose metabolism, intersecting with oxidative stress and carbonyl detoxification processes. AKR1B1 activity is frequently studied in the context of hyperglycemia-associated metabolic stress, inflammation-related signaling, and tissue susceptibility to oxidative damage. Dysregulated polyol flux and altered aldehyde handling have been linked to mechanisms underlying diabetic complications and broader metabolic disease phenotypes in experimental models.
Aldose Reductase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the AKR1B1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within AKR1B1. 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 AKR1B1 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 AKR1B1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.