KCNT1 CRISPR Activation Plasmid (m): sc-432714-ACT

Kcnt1 encodes the mouse KCNT1 sodium-activated potassium channel, a large-conductance K+ channel that couples intracellular Na+ elevations to membrane repolarization and firing-rate adaptation in neurons. By shaping action potential afterhyperpolarization and burst dynamics, KCNT1 influences synaptic integration and network excitability, intersecting with pathways governing ion homeostasis and activity-dependent signaling. Altered KCNT1 activity is linked to neuronal hyperexcitability phenotypes and has been studied in the context of seizure-related and neurodevelopmental disorder mechanisms, supporting its relevance in circuit-level dysfunction. In mouse models, Kcnt1 serves as a tractable node for probing how KNa conductances tune excitatory–inhibitory balance and plasticity.

KCNT1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Kcnt1 expression without altering the underlying DNA sequence.

KCNT1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Kcnt1 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 Kcnt1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KCNT1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Kcnt1 locus and enabling the study of KCNT1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KCNT1 pathway restoration in tumor cells with silenced or reduced Kcnt1 expression.

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