HYAL2 CRISPR Activation Plasmid (h): sc-405005-ACT

Human HYAL2 encodes a glycosylphosphatidylinositol (GPI)-anchored hyaluronidase that participates in extracellular matrix remodeling by initiating hyaluronan (HA) depolymerization at the cell surface. By generating intermediate HA fragments that can be further processed by other hyaluronidases, HYAL2 influences pericellular matrix organization, cell adhesion, and motility, and can modulate receptor-mediated signaling linked to HA biology such as CD44- and RHAMM-associated pathways. Altered HA turnover and HYAL2 activity have been associated with changes in inflammation and tissue fibrosis, and with tumor microenvironment phenotypes including invasion and stromal remodeling. As a result, HYAL2 is frequently studied in contexts where matrix dynamics shape cell behavior, including developmental processes, wound responses, and cancer-associated extracellular matrix regulation.

HYAL2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous HYAL2 expression without altering the underlying DNA sequence.

HYAL2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the HYAL2 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 HYAL2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HYAL2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native HYAL2 locus and enabling the study of HYAL2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HYAL2 pathway restoration in tumor cells with silenced or reduced HYAL2 expression.

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