PMCA1 Double Nickase Plasmid (h): sc-402691-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PMCA1 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
- PMCA1 Double Nickase Plasmid (h) and PMCA1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP2B1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PMCA1 Antibody (F-10): sc-398413
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PMCA1 Double Nickase Plasmid (h) | sc-402691-NIC | 20 µg | $410.00 | |||
PMCA1 Double Nickase Plasmid (h2) | sc-402691-NIC-2 | 20 µg | $410.00 |
ATP2B1 encodes plasma membrane Ca2+-transporting ATPase 1 (PMCA1), a high-affinity Ca2+ efflux pump that maintains low cytosolic calcium and shapes calcium-dependent signaling dynamics. PMCA1 couples ATP hydrolysis to Ca2+ extrusion, supporting processes such as excitability, secretion, cell-cycle progression, and calcium-dependent transcription through pathways linked to calmodulin, calcineurin/NFAT, and CaMK signaling. By regulating local Ca2+ microdomains at the plasma membrane, PMCA1 influences endothelial and smooth muscle function, neuronal homeostasis, and barrier integrity. Genetic and functional perturbation of ATP2B1 has been associated with cardiovascular and neurovascular phenotypes, including blood pressure regulation and calcium-handling defects that are relevant to mechanistic disease modeling in human cells.
PMCA1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP2B1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP2B1. 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 ATP2B1 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 ATP2B1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.