CaM II CRISPR Activation Plasmid (h): sc-400952-ACT

Human CALM2 encodes calmodulin (CaM II), a ubiquitous Ca2+ sensor that transduces intracellular calcium transients into regulatory signals by binding and modulating numerous enzymes, ion channels, and transcriptional regulators. Through Ca2+/calmodulin-dependent signaling, it contributes to processes including excitation–contraction coupling, synaptic plasticity, cell-cycle control, and stimulus-dependent gene expression, intersecting with pathways such as CaMK signaling, calcineurin/NFAT activation, and GPCR-initiated Ca2+ dynamics. CALM2-mediated buffering and decoding of Ca2+ signals supports mitochondrial function and membrane excitability, shaping downstream phosphorylation networks and calcium homeostasis. Dysregulated calmodulin signaling and pathogenic CALM2 variants have been linked to arrhythmogenic phenotypes and broader neurocardiac dysfunction, making CALM2 a relevant target for mechanistic studies of calcium-driven disease biology.

CaM II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CALM2 expression without altering the underlying DNA sequence.

CaM II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CALM2 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 CALM2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaM II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CALM2 locus and enabling the study of CaM II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaM II pathway restoration in tumor cells with silenced or reduced CALM2 expression.

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