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.
Leukocyte-type 12-LO Double Nickase Plasmid (m): sc-419089-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Leukocyte-type 12-LO Double Nickase Plasmid (m) 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
- Leukocyte-type 12-LO Double Nickase Plasmid (m) and Leukocyte-type 12-LO Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Alox15. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: 15-LO Antibody (B-7): sc-133085
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Leukocyte-type 12-LO Double Nickase Plasmid (m) | sc-419089-NIC | 20 µg | $410.00 | |||
Leukocyte-type 12-LO Double Nickase Plasmid (m2) | sc-419089-NIC-2 | 20 µg | $410.00 |
Mouse Alox15 encodes leukocyte-type 12-lipoxygenase (12-LO), a non-heme iron dioxygenase that oxygenates polyunsaturated fatty acids such as arachidonic acid to generate bioactive lipid mediators including 12-HETE and related oxylipins. These products participate in redox-sensitive signaling, regulation of inflammatory responses, leukocyte chemotaxis, and lipid remodeling, linking Alox15 activity to eicosanoid metabolism and resolution programs. In immune and vascular cell contexts, 12-LO-derived metabolites can influence cytokine signaling, oxidative stress, and cell fate decisions that shape tissue inflammation. Consequently, Alox15 is widely studied in models of inflammation, atherosclerosis, metabolic dysfunction, and immune-mediated tissue injury where lipid mediator balance is a key determinant of phenotype.
Leukocyte-type 12-LO Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Alox15 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Alox15. 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 Alox15 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 Alox15-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.