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.
eNOS Double Nickase Plasmid (m): sc-421929-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- eNOS 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
- eNOS Double Nickase Plasmid (m) and eNOS Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Nos3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: eNOS Antibody (A-9): sc-376751
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
eNOS Double Nickase Plasmid (m) | sc-421929-NIC | 20 µg | $410.00 |
Mouse Nos3 encodes endothelial nitric oxide synthase (eNOS), a calcium/calmodulin-regulated enzyme that generates nitric oxide from L-arginine to control vascular tone, platelet–endothelium interactions, and endothelial barrier function. eNOS activity is tuned by phosphorylation and protein–protein interactions downstream of PI3K–AKT, VEGF, and shear-stress signaling, coupling metabolic and inflammatory cues to nitric oxide bioavailability. Dysregulation of Nos3-dependent signaling is broadly implicated in endothelial dysfunction and altered redox balance, with relevance to hypertension, atherosclerosis, ischemia-reperfusion injury, and inflammatory vascular remodeling. In biomedical research, Nos3 serves as a central node for studying nitric oxide–cGMP signaling, oxidative stress, and vascular cell communication in vitro and in vivo.
eNOS Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Nos3 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Nos3. 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 Nos3 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 Nos3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.