PMCA2 CRISPR Activation Plasmid (h): sc-403416-ACT

ATP2B2 encodes the plasma membrane Ca2+ ATPase isoform PMCA2, a high-affinity Ca2+ extrusion pump that maintains low cytosolic calcium and shapes stimulus-dependent Ca2+ transients. By coupling ATP hydrolysis to Ca2+ efflux, PMCA2 supports calcium-dependent signaling networks that regulate neuronal excitability, synaptic transmission, and epithelial transport processes. PMCA2 activity intersects with calmodulin-regulated pathways, membrane microdomain signaling, and homeostatic feedback that prevents Ca2+-driven proteotoxic and mitochondrial stress. Altered ATP2B2/PMCA2 expression or function has been associated with dysregulated calcium handling implicated in neurological phenotypes and sensory system dysfunction, supporting its relevance for mechanistic studies of Ca2+ signaling.

PMCA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATP2B2 expression without altering the underlying DNA sequence.

PMCA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATP2B2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the ATP2B2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PMCA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATP2B2 locus and enabling the study of PMCA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PMCA2 pathway restoration in tumor cells with silenced or reduced ATP2B2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.