KIR7.1 CRISPR Activation Plasmid (h): sc-403910-ACT

KCNJ13 encodes the inwardly rectifying potassium channel KIR7.1, a key regulator of membrane potential and K+ homeostasis in polarized epithelia and excitable tissues. By shaping potassium conductance, KIR7.1 influences ion-coupled transport, epithelial barrier physiology, and electrical signaling processes that intersect with transepithelial fluid movement and retinal pigment epithelium function. Altered KCNJ13 activity has been linked to retinal dysfunction and inherited ocular disease phenotypes, reflecting the channel’s role in maintaining ionic balance in visual pathways. As a membrane channel with tissue-specific expression, KIR7.1 provides a tractable node for dissecting how potassium flux modulates cellular excitability, transport networks, and stress responses.

KIR7.1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KCNJ13 expression without altering the underlying DNA sequence.

KIR7.1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KCNJ13 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 KCNJ13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous KIR7.1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KCNJ13 locus and enabling the study of KIR7.1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of KIR7.1 pathway restoration in tumor cells with silenced or reduced KCNJ13 expression.

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