βENaC CRISPR Activation Plasmid (h): sc-401371-ACT

SCNN1B encodes the β subunit of the epithelial sodium channel (βENaC), a core component of amiloride-sensitive Na⁺ transport across apical membranes in epithelial tissues. In concert with α and γ subunits, βENaC supports sodium reabsorption and fluid homeostasis, linking ion transport to epithelial barrier function and volume regulation pathways. ENaC activity is integrated with proteolytic processing, ubiquitin-dependent trafficking (including NEDD4L-mediated regulation), and hormone-responsive signaling that tunes channel abundance and open probability. Altered SCNN1B expression or function has been associated with disorders of salt handling and airway surface liquid regulation, making it relevant for studies of epithelial physiology and disease-associated ion transport phenotypes.

βENaC CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCNN1B expression without altering the underlying DNA sequence.

βENaC CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCNN1B 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 SCNN1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous βENaC expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCNN1B locus and enabling the study of βENaC-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of βENaC pathway restoration in tumor cells with silenced or reduced SCNN1B expression.

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