SERCA3 CRISPR Activation Plasmid (h): sc-402840-ACT

ATP2A3 encodes the human sarco/endoplasmic reticulum Ca2+-ATPase SERCA3, an ATP-driven pump that transports cytosolic Ca2+ into the endoplasmic reticulum to restore basal calcium levels after signaling events. By shaping ER Ca2+ stores and cytosolic Ca2+ transients, SERCA3 influences calcium-dependent processes including secretion, receptor signaling, transcriptional programs, and ER stress responses. SERCA3 activity contributes to intracellular calcium homeostasis that interfaces with apoptosis regulation, unfolded protein response signaling, and broader Ca2+-regulated kinase and phosphatase pathways. Altered expression of SERCA family members has been associated with dysregulated calcium signaling observed across multiple disease contexts, making ATP2A3 a useful target for mechanistic studies in cell physiology and stress adaptation.

SERCA3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATP2A3 expression without altering the underlying DNA sequence.

SERCA3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATP2A3 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 ATP2A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SERCA3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATP2A3 locus and enabling the study of SERCA3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SERCA3 pathway restoration in tumor cells with silenced or reduced ATP2A3 expression.

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