ADAMTS-9 CRISPR Activation Plasmid (h): sc-402605-ACT

ADAMTS9 encodes ADAMTS-9, a secreted zinc-dependent metalloproteinase that participates in extracellular matrix remodeling through proteolytic processing of proteoglycans such as aggrecan and versican. By regulating matrix composition, pericellular proteolysis, and tissue morphogenesis, ADAMTS-9 influences cell adhesion, migration, and signaling crosstalk with pathways linked to development and inflammation. Altered ADAMTS9 expression or activity has been associated with connective tissue and cartilage-related biology and has been investigated in contexts of tumor microenvironment remodeling and vascular or metabolic phenotypes. These features make ADAMTS-9 a relevant target for mechanistic studies of ECM dynamics, cell–matrix interactions, and protease-regulated signaling.

ADAMTS-9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADAMTS9 expression without altering the underlying DNA sequence.

ADAMTS-9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADAMTS9 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 ADAMTS9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADAMTS-9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADAMTS9 locus and enabling the study of ADAMTS-9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADAMTS-9 pathway restoration in tumor cells with silenced or reduced ADAMTS9 expression.

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