SphK2 Double Nickase Plasmid (h): sc-417921-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SphK2 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
- SphK2 Double Nickase Plasmid (h) and SphK2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting SPHK2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: SphK2 Antibody (9C5E1): sc-517192
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SphK2 Double Nickase Plasmid (h) | sc-417921-NIC | 20 µg | $410.00 | |||
SphK2 Double Nickase Plasmid (h2) | sc-417921-NIC-2 | 20 µg | $410.00 |
SPHK2 encodes sphingosine kinase 2 (SphK2), an enzyme that phosphorylates sphingosine to generate sphingosine-1-phosphate (S1P), a bioactive lipid mediator that integrates metabolic and signaling networks. Through S1P production and subcellular compartmentalization, SphK2 influences MAPK/ERK, PI3K–AKT, and NF-κB-linked processes that shape proliferation, survival, stress responses, and inflammatory signaling. SphK2 activity also interfaces with mitochondrial and nuclear functions, contributing to regulation of apoptosis, epigenetic control, and cellular bioenergetics. Dysregulation of S1P metabolism and SPHK2-associated signaling has been implicated in contexts such as cancer biology, immune dysregulation, and cardiometabolic and neuroinflammatory disease mechanisms.
SphK2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the SPHK2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within SPHK2. 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 SPHK2 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 SPHK2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.