Hepsin CRISPR Activation Plasmid (h): sc-405081-ACT

Human HPN encodes hepsin, a type II transmembrane serine protease localized at the cell surface where it regulates pericellular proteolysis and extracellular matrix remodeling. Hepsin participates in protease-driven signaling by cleaving substrates that can modulate growth factor availability, cell adhesion, and invasive behavior, linking it to pathways governing epithelial organization and tissue homeostasis. Dysregulated HPN expression has been associated with altered epithelial differentiation and tumor-associated proteolytic programs, making it relevant for studying mechanisms of cell migration, barrier integrity, and microenvironmental crosstalk. As a membrane-anchored protease, hepsin is also informative for investigating protease networks and downstream transcriptional responses triggered by surface proteolysis.

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

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

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