CALHM1 CRISPR Activation Plasmid (h): sc-407600-ACT

CALHM1 (calcium homeostasis modulator 1) encodes a plasma membrane ion channel that contributes to calcium-permeable conductance and ATP release, linking extracellular cues to intracellular Ca2+ dynamics and purinergic signaling. CALHM1 activity influences neuronal excitability, synaptic function, and stimulus-secretion coupling by shaping membrane potential and Ca2+-dependent signaling cascades. In the central nervous system, CALHM1 has been studied in the context of amyloid-related pathways and neuroinflammatory signaling, supporting its relevance to mechanistic research in neurodegeneration and cognitive phenotypes. Its channel-mediated regulation of ATP efflux also makes it a useful target for dissecting intercellular communication in excitable and epithelial tissues.

CALHM1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CALHM1 expression without altering the underlying DNA sequence.

CALHM1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CALHM1 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 CALHM1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CALHM1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CALHM1 locus and enabling the study of CALHM1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CALHM1 pathway restoration in tumor cells with silenced or reduced CALHM1 expression.

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