γENaC CRISPR Activation Plasmid (h): sc-402577-ACT

SCNN1G encodes the γ subunit of the epithelial sodium channel (γENaC), a heterotrimeric amiloride-sensitive channel that mediates electrogenic Na⁺ entry across apical membranes in kidney, lung, colon, and other epithelia. γENaC function integrates with aldosterone and vasopressin signaling to regulate transepithelial salt and water transport, influencing extracellular fluid volume, airway surface liquid homeostasis, and systemic electrolyte balance. Channel activity is controlled by proteolytic processing, ubiquitin-dependent trafficking via NEDD4-2, and cellular pathways governing membrane residency and open probability. Dysregulation of ENaC subunits, including SCNN1G, is implicated in disorders of salt handling and airway hydration, supporting mechanistic studies in epithelial physiology and ion transport biology.

γENaC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCNN1G expression without altering the underlying DNA sequence.

γENaC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCNN1G 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 SCNN1G transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous γENaC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCNN1G locus and enabling the study of γENaC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of γENaC pathway restoration in tumor cells with silenced or reduced SCNN1G expression.

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