CaMKII delta CRISPR Activation Plasmid (h): sc-400811-ACT

CAMK2D encodes the delta isoform of Ca2+/calmodulin-dependent protein kinase II (CaMKII delta), a serine/threonine kinase that decodes intracellular calcium transients into phosphorylation-dependent signaling outputs. CaMKII delta regulates excitation–contraction coupling, calcium handling, and activity-dependent transcriptional programs through targets spanning ion channels, sarcoplasmic reticulum proteins, and downstream effectors including MAPK and CREB-associated networks. In human tissues, CAMK2D activity is closely linked to cardiac and vascular signaling, where altered CaMKII delta regulation can reshape stress-responsive pathways, electrophysiology, and cellular remodeling. Dysregulated calcium signaling and CaMKII-dependent phosphorylation events have been associated with cardiovascular pathology and broader calcium-dependent signaling defects relevant to mechanistic disease studies.

CaMKII delta CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAMK2D expression without altering the underlying DNA sequence.

CaMKII delta CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAMK2D 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 CAMK2D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaMKII delta expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAMK2D locus and enabling the study of CaMKII delta-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaMKII delta pathway restoration in tumor cells with silenced or reduced CAMK2D expression.

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