versican CRISPR Activation Plasmid (h): sc-401173-ACT

VCAN encodes versican, a large extracellular matrix chondroitin sulfate proteoglycan that modulates tissue architecture, hydration, and pericellular signaling through interactions with hyaluronan, link proteins, and growth factors. Versican influences cell adhesion, migration, and proliferation by shaping integrin-mediated cues and extracellular matrix remodeling programs, with context-dependent effects on inflammatory signaling and stromal–epithelial crosstalk. Altered VCAN expression and isoform balance have been associated with fibrosis, vascular remodeling, and tumor-associated stroma, where extracellular matrix composition can regulate invasion and immune cell trafficking. These features make VCAN a useful target for studying matrix-driven regulation of cell behavior in development, wound repair, and disease-relevant microenvironments.

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

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

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