VPAC2 CRISPR Activation Plasmid (h): sc-402062-ACT

Human VIPR2 encodes the VPAC2 receptor, a class B G protein–coupled receptor for vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase–activating polypeptide (PACAP). VPAC2 primarily couples to Gs to elevate cAMP and activate PKA-dependent transcriptional programs, with additional effects on MAPK/ERK signaling and calcium dynamics depending on cellular context. This receptor modulates neuroendocrine communication, circadian and metabolic regulation, and immune cell activity by shaping cytokine signaling and cellular responsiveness to neuropeptides. Dysregulated VIPR2/VPAC2 signaling and copy-number variation have been linked to neuropsychiatric and developmental phenotypes, and altered pathway activity has been studied in inflammatory and metabolic disease biology.

VPAC2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VIPR2 expression without altering the underlying DNA sequence.

VPAC2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VIPR2 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 VIPR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VPAC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VIPR2 locus and enabling the study of VPAC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VPAC2 pathway restoration in tumor cells with silenced or reduced VIPR2 expression.

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