ARG-Vasopressin CRISPR Activation Plasmid (h): sc-403139-ACT

AVP encodes arginine vasopressin, a neurohypophyseal peptide hormone synthesized as a preprohormone and processed to regulate systemic water balance, vascular tone, and neuroendocrine homeostasis. Through AVPR2 signaling in renal collecting duct principal cells, vasopressin drives cAMP/PKA-dependent trafficking of aquaporin-2 to the apical membrane, coordinating antidiuretic responses; AVPR1A-mediated pathways contribute to vasoconstriction and broader stress-responsive signaling. AVP-linked dysregulation is relevant to disorders of osmoregulation and blood pressure control, including central diabetes insipidus and inappropriate antidiuresis, and it is also studied in neuropsychiatric and behavioral contexts due to CNS expression patterns. As a tightly regulated hypothalamic gene, AVP provides a model for stimulus-dependent transcription, peptide processing, and secretory pathway biology.

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

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

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