uPA/urokinase/PLAU CRISPR Activation Plasmid (h): sc-400694-ACT

PLAU encodes urokinase-type plasminogen activator (uPA), a secreted serine protease that converts plasminogen to plasmin to promote pericellular proteolysis and extracellular matrix remodeling. Through interactions with the uPA receptor (PLAUR/uPAR), uPA concentrates proteolytic activity at the cell surface and engages signaling networks that influence adhesion and migration, including integrin- and growth factor–linked pathways. PLAU activity shapes fibrinolysis, wound repair, and tissue remodeling, and its dysregulation is frequently studied in contexts of invasive cell behavior and inflammatory microenvironments. As a result, PLAU is widely used as a functional node for investigating protease-driven matrix dynamics, cell motility, and remodeling-associated gene programs.

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

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

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