SERCA2 CRISPR Activation Plasmid (h): sc-400417-ACT

ATP2A2 encodes the human SERCA2 sarco/endoplasmic reticulum Ca²⁺-ATPase, a P-type ATPase that pumps cytosolic Ca²⁺ into the ER/SR lumen to sustain calcium homeostasis and refill intracellular stores. By shaping Ca²⁺ transients, SERCA2 influences excitation–contraction coupling, ER protein folding capacity, and calcium-dependent signaling networks linked to stress responses and apoptosis. SERCA2 activity is integrated with phospholamban-mediated regulation and broader ion transport pathways that govern cellular bioenergetics and contractility. Dysregulation or mutation of ATP2A2 is associated with altered ER Ca²⁺ handling and has been implicated in disorders of epithelial integrity and cardiac physiology, providing a mechanistic entry point for studying calcium-dependent disease phenotypes.

SERCA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATP2A2 expression without altering the underlying DNA sequence.

SERCA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATP2A2 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 ATP2A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SERCA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATP2A2 locus and enabling the study of SERCA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SERCA2 pathway restoration in tumor cells with silenced or reduced ATP2A2 expression.

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