VIP CRISPR Activation Plasmid (h): sc-400776-ACT

Vasoactive intestinal peptide (VIP) is a secreted neuropeptide hormone that signals primarily through the class B GPCRs VPAC1 (VIPR1) and VPAC2 (VIPR2) to stimulate adenylate cyclase, elevate cAMP, and activate PKA-dependent transcriptional programs. In humans, VIP contributes to neuroendocrine communication, smooth muscle relaxation, circadian and neuroimmune regulation, and epithelial secretion, with downstream effects on MAPK signaling and cytokine networks. VIP pathway activity is frequently studied in contexts of inflammation and immune cell polarization, airway and gastrointestinal physiology, and neurobiology, where altered VIP signaling has been associated with tumor microenvironment modulation and neuroinflammatory states. Because VIP is rapidly processed and acts in a paracrine/endocrine manner, controlled upregulation is useful for dissecting receptor-specific signaling dynamics and secreted peptide biology.

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

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

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