KCNQ4 CRISPR Activation Plasmid (h): sc-403090-ACT

KCNQ4 encodes a voltage-gated potassium channel subunit that contributes to M-type K+ currents and helps set resting membrane potential and cellular excitability. By regulating membrane repolarization and potassium ion homeostasis, KCNQ4 influences electrophysiological signaling in excitable tissues, including sensory epithelia. Dysregulated KCNQ4 activity has been linked to auditory dysfunction and progressive sensorineural hearing phenotypes, making it relevant for studies of cochlear hair cell physiology and ion channel–dependent stress responses. KCNQ4 is also used as a mechanistic entry point for investigating channelopathy-associated signaling programs and transcriptional adaptations to altered K+ conductance.

KCNQ4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNQ4 expression without altering the underlying DNA sequence.

KCNQ4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNQ4 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 KCNQ4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KCNQ4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNQ4 locus and enabling the study of KCNQ4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KCNQ4 pathway restoration in tumor cells with silenced or reduced KCNQ4 expression.

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