KV1.5 CRISPR Activation Plasmid (h): sc-404341-ACT

KCNA5 encodes the human voltage-gated potassium channel KV1.5, a Shaker-related α-subunit that conducts the ultrarapid delayed rectifier current and helps control membrane repolarization and excitability. KV1.5 activity shapes action potential duration, firing frequency, and calcium-coupled signaling by tuning resting membrane potential and refractory behavior in excitable and non-excitable cells. Through its contributions to electrophysiological homeostasis, KCNA5 is frequently studied in pathways linking ion flux to transcriptional programs, metabolic demand, and stress responses. Dysregulated KCNA5/KV1.5 expression or function has been associated with cardiac rhythm phenotypes, vascular smooth muscle remodeling, and altered proliferative signaling, supporting its relevance in mechanistic models of channelopathies.

KV1.5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNA5 expression without altering the underlying DNA sequence.

KV1.5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNA5 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 KCNA5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KV1.5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNA5 locus and enabling the study of KV1.5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KV1.5 pathway restoration in tumor cells with silenced or reduced KCNA5 expression.

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