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.
N-SMase2 Double Nickase Plasmid (h): sc-401937-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- N-SMase2 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
- N-SMase2 Double Nickase Plasmid (h) and N-SMase2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SMPD3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: N-SMase2 Antibody (G-6): sc-166637
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
N-SMase2 Double Nickase Plasmid (h) | sc-401937-NIC | 20 µg | $410.00 | |||
N-SMase2 Double Nickase Plasmid (h2) | sc-401937-NIC-2 | 20 µg | $410.00 |
Human SMPD3 encodes neutral sphingomyelinase 2 (N‑SMase2), a membrane-associated enzyme that hydrolyzes sphingomyelin to generate ceramide and phosphocholine. By controlling ceramide production at the plasma membrane and Golgi, N‑SMase2 regulates sphingolipid metabolism, membrane microdomain organization, and signal transduction downstream of cellular stress and inflammatory cues. SMPD3 activity has been linked to pathways influencing apoptosis, vesicle trafficking, and extracellular vesicle biogenesis, with downstream effects on lipid homeostasis and cell-state regulation. Dysregulated ceramide signaling and altered SMPD3/N‑SMase2 function are implicated in disease-relevant processes including neurobiology, metabolic dysfunction, inflammation, and cancer-associated phenotypes.
N-SMase2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SMPD3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SMPD3. 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 SMPD3 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 SMPD3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.