KV4.3 CRISPR Activation Plasmid (h): sc-404247-ACT

KCND3 encodes the human voltage-gated potassium channel subunit KV4.3, a principal mediator of fast, transient A-type potassium currents that shape action potential repolarization and firing frequency. KV4.3 function integrates with membrane excitability networks in neurons and cardiomyocytes, influencing synaptic integration, conduction dynamics, and activity-dependent signaling. By tuning repolarization kinetics and excitability thresholds, KCND3 intersects with pathways governing electrical signaling, calcium coupling, and downstream transcriptional programs responsive to depolarization. Genetic and functional perturbations in KCND3 have been linked to disorders of excitability, including neurological phenotypes such as spinocerebellar ataxia and cardiac electrical abnormalities, supporting its relevance in mechanism-focused studies.

KV4.3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCND3 expression without altering the underlying DNA sequence.

KV4.3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCND3 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 KCND3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KV4.3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCND3 locus and enabling the study of KV4.3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KV4.3 pathway restoration in tumor cells with silenced or reduced KCND3 expression.

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