PMCA3 Double Nickase Plasmid (h): sc-404464-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PMCA3 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
- PMCA3 Double Nickase Plasmid (h) and PMCA3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP2B3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PMCA3 Antibody (G-6): sc-390148
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PMCA3 Double Nickase Plasmid (h) | sc-404464-NIC | 20 µg | $410.00 | |||
PMCA3 Double Nickase Plasmid (h2) | sc-404464-NIC-2 | 20 µg | $410.00 |
Human ATP2B3 encodes plasma membrane Ca2+-transporting ATPase 3 (PMCA3), a P-type ATPase that exports cytosolic Ca2+ to maintain low basal calcium and shape stimulus-evoked Ca2+ transients. By restoring Ca2+ homeostasis after signaling events, PMCA3 helps regulate calcium-dependent processes including excitability, secretion, transcriptional responses, and cell survival pathways. PMCA3 activity intersects functionally with calmodulin regulation and broader Ca2+ signaling networks such as calcineurin/NFAT and CaMK-mediated pathways. Genetic disruption or altered expression of ATP2B3 has been associated with neurodevelopmental and neurological phenotypes, supporting its relevance for mechanistic studies of calcium-handling defects.
PMCA3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP2B3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP2B3. 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 ATP2B3 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 ATP2B3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.