CAMK2A/CaMKII alpha CRISPR Activation Plasmid (h): sc-400228-ACT

CAMK2A encodes the alpha subunit of Ca2+/calmodulin-dependent protein kinase II (CaMKIIα), a serine/threonine kinase that decodes intracellular calcium transients into phosphorylation-dependent signaling. In neurons, CaMKIIα is highly enriched at postsynaptic densities where it regulates synaptic vesicle cycling, AMPA receptor trafficking, and activity-dependent gene expression that underpins synaptic plasticity, learning, and memory. CAMK2A integrates upstream inputs from NMDA receptor–mediated Ca2+ influx and intersects with MAPK/ERK and CREB-associated transcriptional programs. Dysregulated CaMKIIα signaling has been implicated in neurodevelopmental and neuropsychiatric phenotypes, neurodegenerative processes, and excitotoxic stress pathways, making it a widely used node for mechanistic studies of neuronal function.

CAMK2A/CaMKII alpha CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAMK2A expression without altering the underlying DNA sequence.

CAMK2A/CaMKII alpha CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAMK2A 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 CAMK2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CAMK2A/CaMKII alpha expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAMK2A locus and enabling the study of CAMK2A/CaMKII alpha-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CAMK2A/CaMKII alpha pathway restoration in tumor cells with silenced or reduced CAMK2A expression.

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