MMP9 CRISPR Activation Plasmid (h): sc-400083-ACT

Human MMP9 encodes matrix metalloproteinase-9, a secreted zinc-dependent endopeptidase that degrades extracellular matrix components such as type IV collagen and regulates basement membrane remodeling. Its activity influences cell migration, angiogenesis, and leukocyte trafficking by shaping the pericellular proteolytic environment and releasing or activating matrix-bound growth factors and cytokines. MMP9 is integrated with inflammatory signaling networks and extracellular matrix organization programs, and it intersects with pathways controlling epithelial–mesenchymal dynamics and tissue repair. Dysregulated MMP9 expression and activity are frequently studied in cancer invasion and metastasis, chronic inflammatory disorders, cardiovascular remodeling, and neuroinflammation.

MMP9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MMP9 expression without altering the underlying DNA sequence.

MMP9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MMP9 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 MMP9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MMP9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MMP9 locus and enabling the study of MMP9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MMP9 pathway restoration in tumor cells with silenced or reduced MMP9 expression.

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