CaMKIV CRISPR Activation Plasmid (h): sc-400806-ACT

Human CAMK4 encodes CaMKIV, a nuclear Ca2+/calmodulin-dependent serine/threonine kinase that couples calcium transients to transcriptional programs controlling neuronal plasticity, immune cell activation, and cell-cycle–linked gene expression. CaMKIV is activated downstream of CaMKK and integrates signals from calcium influx and GPCR pathways to phosphorylate transcriptional regulators such as CREB and modulate chromatin-associated processes. Through these mechanisms it influences differentiation and stimulus-dependent gene induction in neurons and lymphocytes, with reported associations to dysregulated inflammatory signaling and neuropsychiatric phenotypes. Its pathway positioning makes CAMK4 a useful node for dissecting calcium-dependent transcription, synaptic signaling, and immune regulatory networks.

CaMKIV CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAMK4 expression without altering the underlying DNA sequence.

CaMKIV CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAMK4 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 CAMK4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaMKIV expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAMK4 locus and enabling the study of CaMKIV-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaMKIV pathway restoration in tumor cells with silenced or reduced CAMK4 expression.

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