12-LO Double Nickase Plasmid (h): sc-403010-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- 12-LO 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
- 12-LO Double Nickase Plasmid (h) and 12-LO Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ALOX12. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: 12-LO Antibody (C-5): sc-365194
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
12-LO Double Nickase Plasmid (h) | sc-403010-NIC | 20 µg | $410.00 | |||
12-LO Double Nickase Plasmid (h2) | sc-403010-NIC-2 | 20 µg | $410.00 |
Human ALOX12 encodes 12-lipoxygenase (12-LO), a non-heme iron dioxygenase that catalyzes oxygenation of arachidonic acid to generate 12-hydroperoxyeicosatetraenoic acid and downstream lipid mediators that shape inflammatory signaling. 12-LO-derived oxylipins influence platelet activation and aggregation, vascular and epithelial responses, and redox-sensitive processes through modulation of MAPK, NF-κB, and related eicosanoid networks. ALOX12 activity intersects with lipid peroxidation and membrane remodeling pathways, impacting cell migration, differentiation, and responses to oxidative stress. Dysregulated ALOX12/12-LO signaling has been associated with pro-inflammatory states and metabolic and cardiovascular phenotypes, supporting its study in disease-relevant cellular models.
12-LO Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ALOX12 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ALOX12. 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 ALOX12 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 ALOX12-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.