AVP Receptor V2 CRISPR Activation Plasmid (h): sc-403177-ACT

AVPR2 encodes the human arginine vasopressin receptor 2 (AVP Receptor V2), a G protein–coupled receptor predominantly coupled to Gs that activates adenylyl cyclase and elevates intracellular cAMP. Signaling through cAMP/PKA regulates phosphorylation and trafficking programs that influence epithelial water permeability and membrane protein localization, integrating with broader GPCR desensitization and endocytic recycling processes. AVPR2 activity is central to vasopressin-responsive pathways and serves as a model system for studying receptor signaling dynamics, second-messenger regulation, and ligand-dependent transcriptional responses. Genetic variation or dysregulated expression of AVPR2 has been associated with altered water-balance phenotypes and receptor misfolding/trafficking defects, providing mechanistic links to disorders of renal concentrating function.

AVP Receptor V2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous AVPR2 expression without altering the underlying DNA sequence.

AVP Receptor V2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the AVPR2 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 AVPR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AVP Receptor V2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native AVPR2 locus and enabling the study of AVP Receptor V2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AVP Receptor V2 pathway restoration in tumor cells with silenced or reduced AVPR2 expression.

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