KV1.3 CRISPR Activation Plasmid (h): sc-403814-ACT

KCNA3 encodes the human voltage-gated potassium channel KV1.3, a membrane conductance that helps set resting membrane potential and shapes action potential firing in excitable and non-excitable cells. By regulating potassium efflux, KV1.3 influences calcium-dependent signaling programs that control activation, proliferation, and cytokine production, linking ion homeostasis to downstream transcriptional responses. KV1.3 activity intersects with immune cell receptor signaling, mitochondrial bioenergetics, and redox-sensitive pathways that tune cellular activation thresholds. Dysregulated KCNA3/KV1.3 expression or function has been implicated in immune-mediated inflammation and altered proliferative states, supporting its relevance for mechanistic studies in immunology and disease-associated cellular phenotypes.

KV1.3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNA3 expression without altering the underlying DNA sequence.

KV1.3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNA3 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 KCNA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KV1.3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNA3 locus and enabling the study of KV1.3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KV1.3 pathway restoration in tumor cells with silenced or reduced KCNA3 expression.

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