caveolin-3 CRISPR Activation Plasmid (h): sc-400569-ACT

Human CAV3 encodes caveolin-3, a muscle-enriched structural component of caveolae that organizes membrane microdomains and scaffolds signaling proteins at the sarcolemma. Caveolin-3 influences mechanotransduction, lipid homeostasis, and receptor-associated pathways by modulating compartmentalization of ion channels and kinases involved in muscle excitability and metabolism. Perturbation of CAV3 expression or caveolar architecture is linked to skeletal and cardiac muscle dysfunction, including forms of muscular dystrophy and cardiomyopathy, supporting its relevance in myofiber stability and stress responses. These properties make caveolin-3 a useful entry point for studying membrane trafficking, signal transduction, and muscle cell differentiation in human model systems.

caveolin-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CAV3 expression without altering the underlying DNA sequence.

caveolin-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CAV3 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 CAV3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous caveolin-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CAV3 locus and enabling the study of caveolin-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of caveolin-3 pathway restoration in tumor cells with silenced or reduced CAV3 expression.

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