KIR3.2 CRISPR Activation Plasmid (h): sc-405031-ACT

KCNJ6 encodes the human inwardly rectifying potassium channel subunit KIR3.2 (GIRK2), a key effector of G protein–coupled receptor signaling that helps stabilize resting membrane potential and dampen cellular excitability. Upon activation, Gβγ released from Gi/o-coupled receptors promotes channel opening, coupling neurotransmitter and neuromodulator inputs to potassium conductance in neurons and other excitable cells. KIR3.2 contributes to regulation of action potential firing, synaptic integration, and network oscillations, linking it to pathways that shape neurophysiological responses. Dysregulated KCNJ6 activity or expression has been associated with altered neuronal signaling and has been investigated in contexts such as neurodevelopmental and neuropsychiatric phenotypes, as well as seizure susceptibility.

KIR3.2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNJ6 expression without altering the underlying DNA sequence.

KIR3.2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNJ6 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 KCNJ6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIR3.2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNJ6 locus and enabling the study of KIR3.2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIR3.2 pathway restoration in tumor cells with silenced or reduced KCNJ6 expression.

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