Matriptase CRISPR Activation Plasmid (h): sc-402971-ACT

Human ST14 encodes matriptase, a type II transmembrane serine protease that regulates pericellular proteolysis at epithelial surfaces. Matriptase participates in protease cascades by activating substrates such as pro-hepatocyte growth factor and other extracellular or membrane-associated proteins, influencing processes including epithelial barrier homeostasis, cell polarity, and remodeling of the local microenvironment. By shaping extracellular matrix turnover and growth factor availability, ST14 activity can modulate signaling pathways linked to proliferation, differentiation, and migration. Dysregulated matriptase expression or proteolytic activity has been associated with epithelial pathobiology and is frequently studied in cancer biology, inflammation, and tissue remodeling contexts.

Matriptase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ST14 expression without altering the underlying DNA sequence.

Matriptase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ST14 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 ST14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Matriptase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ST14 locus and enabling the study of Matriptase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Matriptase pathway restoration in tumor cells with silenced or reduced ST14 expression.

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