CaMKIδ CRISPR Activation Plasmid (h): sc-403073-ACT

CAMK1D encodes CaMKIδ, a Ca2+/calmodulin-dependent serine/threonine kinase that converts intracellular calcium transients into phosphorylation events shaping gene expression and cell state decisions. CaMKIδ participates in calcium signaling networks that intersect with CREB-linked transcriptional programs, MAPK-related responses, and cytoskeletal dynamics, supporting context-dependent regulation of proliferation, differentiation, and stress signaling. Genetic and functional studies associate CAMK1D with immunometabolic regulation and inflammatory signaling, with reported links to cardiometabolic phenotypes and susceptibility to complex diseases where calcium-dependent signaling is dysregulated. These features make CAMK1D a useful locus for dissecting how calcium-driven kinase activity influences downstream transcriptional outputs across diverse human cell types.

CaMKIδ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAMK1D expression without altering the underlying DNA sequence.

CaMKIδ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAMK1D 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 CAMK1D transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaMKIδ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAMK1D locus and enabling the study of CaMKIδ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaMKIδ pathway restoration in tumor cells with silenced or reduced CAMK1D expression.

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