ADAMTS-5 CRISPR Activation Plasmid (h): sc-402695-ACT

ADAMTS5 encodes ADAMTS-5, a secreted zinc-dependent metalloproteinase that cleaves extracellular matrix proteoglycans, particularly aggrecan and versican, and thereby regulates matrix turnover and tissue remodeling. ADAMTS-5 activity influences cartilage homeostasis, pericellular matrix composition, and cell–matrix signaling events that intersect with inflammatory mediators and growth factor availability. Dysregulated ADAMTS5 expression or proteolytic activity has been linked to accelerated extracellular matrix degradation in degenerative joint processes and broader connective tissue pathology. As a matrix-modifying enzyme, ADAMTS-5 is commonly studied in pathways governing cartilage catabolism, stromal remodeling, and protease–inhibitor balance.

ADAMTS-5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAMTS5 expression without altering the underlying DNA sequence.

ADAMTS-5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAMTS5 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 ADAMTS5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAMTS-5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAMTS5 locus and enabling the study of ADAMTS-5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAMTS-5 pathway restoration in tumor cells with silenced or reduced ADAMTS5 expression.

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