VE-cadherin-2 CRISPR Activation Plasmid (h): sc-401695-ACT

PCDH12 encodes VE-cadherin-2, a calcium-dependent protocadherin enriched at endothelial cell–cell junctions where it supports adhesion, barrier integrity, and coordinated vascular morphogenesis. Through homophilic interactions and coupling to cytoskeletal and signaling adaptors, VE-cadherin-2 influences junctional remodeling, cell polarity, and migration programs that intersect with angiogenic and inflammatory signaling networks. Dysregulated endothelial adhesion and permeability linked to altered PCDH12 activity has been associated with vascular dysfunction phenotypes and neurovascular developmental disorders, making it relevant for studies of vessel stability and tissue perfusion. Because junctional cadherins also modulate leukocyte transmigration and shear-stress responses, PCDH12 is frequently examined in models of endothelial activation and microvascular pathology.

VE-cadherin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PCDH12 expression without altering the underlying DNA sequence.

VE-cadherin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PCDH12 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 PCDH12 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VE-cadherin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PCDH12 locus and enabling the study of VE-cadherin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VE-cadherin-2 pathway restoration in tumor cells with silenced or reduced PCDH12 expression.

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