PMCA4 Double Nickase Plasmid (h): sc-402888-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PMCA4 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
- PMCA4 Double Nickase Plasmid (h) and PMCA4 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP2B4. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PMCA4 Double Nickase Plasmid (h) | sc-402888-NIC | 20 µg | $410.00 | |||
PMCA4 Double Nickase Plasmid (h2) | sc-402888-NIC-2 | 20 µg | $410.00 |
ATP2B4 encodes plasma membrane Ca2+-ATPase 4 (PMCA4), a P-type ATPase that extrudes cytosolic Ca2+ to maintain calcium homeostasis and shape the amplitude and duration of Ca2+-dependent signaling. PMCA4 regulates localized Ca2+ microdomains that influence processes such as excitation–contraction coupling, nitric oxide signaling, and Ca2+-dependent transcriptional programs. By controlling Ca2+ clearance at the plasma membrane, PMCA4 interfaces with pathways including calmodulin-dependent regulation and downstream kinase/phosphatase networks that tune cell motility, secretion, and proliferation. Genetic variation or altered expression of ATP2B4 has been associated with phenotypes linked to erythrocyte biology and vascular function, supporting its relevance in mechanistic studies of calcium-driven cellular physiology.
PMCA4 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP2B4 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP2B4. 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 ATP2B4 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 ATP2B4-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.