Vitronectin CRISPR Activation Plasmid (h): sc-401129-ACT

Human VTN encodes vitronectin, a secreted glycoprotein abundant in plasma and the extracellular matrix that binds integrins and multiple proteases to coordinate cell adhesion, migration, and tissue remodeling. Vitronectin participates in ECM–receptor interaction and focal adhesion signaling, supporting cytoskeletal organization and survival pathways while modulating proteolysis through interactions with plasminogen activator inhibitor-1. It also interfaces with complement regulation and hemostatic processes, influencing inflammatory microenvironments and matrix turnover. Dysregulated VTN expression or altered vitronectin deposition has been associated with fibrosis, atherosclerosis, cancer invasion and metastasis, and neurodegenerative pathology, making it a useful node for studying extracellular signaling and disease-relevant remodeling programs.

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

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

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