CaMKII beta CRISPR Activation Plasmid (h): sc-402744-ACT

CAMK2B encodes the beta subunit of CaMKII, a Ca2+/calmodulin-dependent serine/threonine kinase that integrates intracellular calcium transients into phosphorylation programs controlling synaptic signaling and structural plasticity. CaMKII beta contributes to holoenzyme assembly, subcellular targeting, and actin cytoskeleton interactions that shape dendritic spine remodeling and activity-dependent neurotransmission. In neurons, CAMK2B participates in pathways linked to NMDA receptor signaling, long-term potentiation/long-term depression, and calcium-dependent gene regulation. Dysregulated CaMKII signaling and CAMK2B-associated network changes have been implicated in neurodevelopmental and neuropsychiatric phenotypes, supporting its use as a mechanistic node for studying neuronal circuit function and plasticity.

CaMKII beta CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAMK2B expression without altering the underlying DNA sequence.

CaMKII beta CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAMK2B 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 CAMK2B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaMKII beta expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAMK2B locus and enabling the study of CaMKII beta-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaMKII beta pathway restoration in tumor cells with silenced or reduced CAMK2B expression.

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