KIR2.1 CRISPR Activation Plasmid (h): sc-401974-ACT

KCNJ2 encodes the inwardly rectifying potassium channel subunit KIR2.1, a key determinant of resting membrane potential and action potential repolarization in excitable cells. By conducting IK1 currents, KIR2.1 stabilizes membrane excitability and shapes electrophysiological behavior in cardiac and skeletal muscle, integrating with ion homeostasis pathways that couple membrane voltage to calcium handling and contractility. Dysregulation of KCNJ2 activity or expression is associated with inherited arrhythmia phenotypes and neuromuscular dysfunction, making it a relevant target for mechanistic studies of channelopathies and electrical signaling. In vitro, modulation of KIR2.1 levels is commonly used to probe membrane potential–dependent transcriptional programs, excitability thresholds, and ion channel network interactions.

KIR2.1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNJ2 expression without altering the underlying DNA sequence.

KIR2.1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNJ2 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 KCNJ2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIR2.1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNJ2 locus and enabling the study of KIR2.1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIR2.1 pathway restoration in tumor cells with silenced or reduced KCNJ2 expression.

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