CaM III CRISPR Activation Plasmid (h): sc-400953-ACT

Human CALM3 encodes calmodulin III (CaM III), a ubiquitous Ca2+-binding sensor that transduces intracellular calcium signals into coordinated changes in enzyme activity, ion channel function, and gene expression. Upon Ca2+ binding, CaM III modulates key effectors including CaMKs, calcineurin, and phosphodiesterases, integrating signaling across neuronal excitability, cardiac conduction, secretion, and cytoskeletal remodeling. CALM3-dependent pathways intersect with MAPK/ERK and CREB-driven transcriptional programs, shaping cell-cycle progression, differentiation, and stress responses. Dysregulated Ca2+/calmodulin signaling is implicated in arrhythmia susceptibility, neurodevelopmental phenotypes, and cancer-associated signaling rewiring, supporting its broad relevance in mechanistic studies.

CaM III CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CALM3 expression without altering the underlying DNA sequence.

CaM III CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CALM3 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 CALM3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CaM III expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CALM3 locus and enabling the study of CaM III-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CaM III pathway restoration in tumor cells with silenced or reduced CALM3 expression.

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