Vasorin CRISPR Activation Plasmid (h): sc-404942-ACT

Human VASN encodes vasorin, a type I transmembrane glycoprotein enriched in vascular smooth muscle cells that modulates cell–cell signaling at the plasma membrane. Vasorin can bind and sequester TGF-β family ligands, thereby influencing SMAD-dependent transcriptional programs that govern extracellular matrix remodeling, phenotypic switching, and migration. Through these activities, VASN is studied in the context of vascular development and injury responses, as well as dysregulated remodeling processes linked to fibrosis and cancer-associated stromal signaling. Altered vasorin expression has been reported in multiple disease settings where TGF-β pathway tone and cell surface shedding are perturbed.

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

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

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