Aquaporin 2/AQP2 CRISPR Activation Plasmid (h): sc-400402-ACT

Aquaporin 2 (AQP2) is a vasopressin-regulated water channel predominantly expressed in kidney collecting duct principal cells, where it controls transepithelial water permeability and systemic water homeostasis. AQP2 trafficking to the apical membrane is governed by AVPR2–cAMP–PKA signaling and phosphorylation-dependent interactions that modulate vesicle recycling, membrane insertion, and endocytosis. Through coupling of osmotic water transport to epithelial polarity and tight junction function, AQP2 contributes to renal concentrating mechanisms and adaptation to hydration status. Dysregulated AQP2 expression, phosphorylation, or trafficking is associated with impaired urinary concentration phenotypes, including nephrogenic diabetes insipidus and disorders of water balance linked to collecting duct dysfunction.

Aquaporin 2/AQP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AQP2 expression without altering the underlying DNA sequence.

Aquaporin 2/AQP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AQP2 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 AQP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Aquaporin 2/AQP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AQP2 locus and enabling the study of Aquaporin 2/AQP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Aquaporin 2/AQP2 pathway restoration in tumor cells with silenced or reduced AQP2 expression.

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