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-SMase Double Nickase Plasmid (h): sc-405568-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- N-SMase 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-SMase Double Nickase Plasmid (h) and N-SMase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SMPD2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: N-SMase Antibody (56-7): sc-100593
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
N-SMase Double Nickase Plasmid (h) | sc-405568-NIC | 20 µg | $410.00 | |||
N-SMase Double Nickase Plasmid (h2) | sc-405568-NIC-2 | 20 µg | $410.00 |
Human SMPD2 encodes neutral sphingomyelinase (N-SMase), a membrane-associated enzyme that hydrolyzes sphingomyelin to generate ceramide and phosphocholine, thereby shaping sphingolipid composition and signaling outputs. N-SMase activity contributes to ceramide-dependent regulation of stress responses, membrane microdomain dynamics, endomembrane trafficking, and crosstalk with apoptotic and inflammatory signaling programs. Through control of ceramide flux, SMPD2 influences pathways linked to ER stress and lipid homeostasis, including downstream effects on MAPK/NF-κB signaling and vesicular transport. Dysregulated sphingolipid metabolism involving SMPD2 has been studied in contexts such as neurodegeneration, metabolic dysfunction, and cancer-associated stress adaptation, supporting its relevance as a research target in disease biology.
N-SMase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SMPD2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SMPD2. 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 SMPD2 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 SMPD2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.