uPAR CRISPR Activation Plasmid (h): sc-400666-ACT

PLAUR encodes the human urokinase plasminogen activator receptor (uPAR), a GPI-anchored cell-surface receptor that localizes proteolysis by binding uPA and coordinating plasminogen activation. Through interactions with integrins, vitronectin, and receptor complexes that include LRP1, uPAR modulates extracellular matrix remodeling, cell adhesion, chemotaxis, and migration, linking pericellular protease activity to cytoskeletal and signaling pathways such as MAPK/ERK, PI3K/AKT, and focal adhesion dynamics. Altered PLAUR/uPAR expression is associated with invasive phenotypes and inflammatory remodeling across multiple disease contexts, including cancer progression, fibrosis, and vascular pathology. As a regulator of protease-driven microenvironmental changes, uPAR is widely studied in mechanisms of metastasis, wound repair, and immune cell trafficking.

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

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

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