HPA1 CRISPR Activation Plasmid (h): sc-402215-ACT

Human HPSE encodes heparanase (HPA1), an endo-β-D-glucuronidase that cleaves heparan sulfate chains on proteoglycans, remodeling extracellular matrix and basement membranes. By liberating heparan sulfate–bound growth factors and cytokines, HPA1 modulates cell–matrix interactions, chemokine gradients, angiogenic signaling, and inflammatory cell trafficking, intersecting with pathways controlling adhesion, migration, and tissue permeability. HPSE activity is linked to altered tumor microenvironment dynamics, invasion and metastasis biology, and dysregulated inflammation in multiple disease contexts. As a result, HPSE is widely used as a mechanistic node for studying extracellular matrix turnover, immune cell recruitment, and proteoglycan-mediated signaling.

HPA1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HPSE expression without altering the underlying DNA sequence.

HPA1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HPSE 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 HPSE transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HPA1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HPSE locus and enabling the study of HPA1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HPA1 pathway restoration in tumor cells with silenced or reduced HPSE expression.

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