KIR6.2 CRISPR Activation Plasmid (m): sc-421232-ACT

Mouse Kcnj11 encodes KIR6.2, the pore-forming inwardly rectifying potassium channel subunit that partners with SUR1/ABCC8 to form ATP-sensitive KATP channels. By coupling intracellular ATP/ADP ratios to membrane potential, KIR6.2 regulates excitability and stimulus-secretion coupling, particularly in pancreatic β cells, and also contributes to metabolic sensing in neurons, cardiac myocytes, and skeletal muscle. KATP channel gating influences calcium influx, mitochondrial function, and downstream signaling that coordinates insulin release and cellular responses to energetic stress. Dysregulation of Kcnj11/KIR6.2 activity is broadly relevant to studies of glucose homeostasis, β-cell dysfunction, and channelopathies impacting electrical signaling and metabolism.

KIR6.2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Kcnj11 expression without altering the underlying DNA sequence.

KIR6.2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Kcnj11 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 Kcnj11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIR6.2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Kcnj11 locus and enabling the study of KIR6.2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIR6.2 pathway restoration in tumor cells with silenced or reduced Kcnj11 expression.

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