Aquaporin 5/AQP5 CRISPR Activation Plasmid (h): sc-400567-ACT

AQP5 encodes aquaporin 5, a membrane water channel that facilitates rapid, osmotically driven water transport across the plasma membrane in secretory epithelia. It supports fluid homeostasis and secretion in tissues such as salivary and lacrimal glands and the airway, contributing to epithelial barrier function and regulation of mucosal hydration. AQP5 localization and abundance are dynamically regulated by signaling pathways that control membrane trafficking and phosphorylation-dependent channel availability, linking it to ion and water movement during glandular secretion. Dysregulated AQP5 expression has been associated with altered airway surface liquid balance, inflammatory epithelial remodeling, and glandular dysfunction, motivating mechanistic studies in respiratory and exocrine biology.

Aquaporin 5/AQP5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AQP5 expression without altering the underlying DNA sequence.

Aquaporin 5/AQP5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AQP5 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 AQP5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Aquaporin 5/AQP5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AQP5 locus and enabling the study of Aquaporin 5/AQP5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Aquaporin 5/AQP5 pathway restoration in tumor cells with silenced or reduced AQP5 expression.

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