PLVAP/PV1 CRISPR Activation Plasmid (h): sc-403746-ACT

PLVAP (plasmalemma vesicle–associated protein), also known as PV1, is an endothelial-restricted membrane glycoprotein that organizes diaphragms of fenestrae, caveolae, and transendothelial channels, thereby regulating vascular permeability and macromolecular transport. By shaping endothelial barrier properties and vesicular trafficking, PLVAP interfaces with processes such as angiogenic remodeling, leukocyte extravasation, and tissue-specific microvascular specialization. Altered PLVAP expression is frequently used as a marker of endothelial activation and pathological neovascularization, and has been associated with inflammatory vascular leakage and tumor-associated endothelium. These features make PLVAP a useful node for studying microvascular function, endothelial differentiation states, and permeability-related phenotypes in human cell models.

PLVAP/PV1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLVAP expression without altering the underlying DNA sequence.

PLVAP/PV1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLVAP 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 PLVAP transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLVAP/PV1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLVAP locus and enabling the study of PLVAP/PV1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLVAP/PV1 pathway restoration in tumor cells with silenced or reduced PLVAP expression.

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