MT-MMP-2 CRISPR Activation Plasmid (h): sc-404075-ACT

MMP15 encodes the membrane-anchored matrix metalloproteinase MT-MMP-2 (MT2-MMP), a zinc-dependent endopeptidase that remodels the pericellular extracellular matrix by cleaving collagens, laminins, and additional matrix substrates at the cell surface. As part of protease networks that coordinate focal adhesion turnover, cell migration, and invasive growth, MT2-MMP influences extracellular signaling by releasing or activating matrix-bound factors and modulating receptor availability. MMP15 activity intersects with integrin- and growth factor–linked pathways that shape epithelial–mesenchymal plasticity and tissue remodeling. Dysregulated MT2-MMP expression has been associated with pathological matrix turnover observed in cancer invasion, fibrosis, and inflammatory microenvironments, making it a useful node for studying proteolysis-driven remodeling programs.

MT-MMP-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MMP15 expression without altering the underlying DNA sequence.

MT-MMP-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MMP15 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 MMP15 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MT-MMP-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MMP15 locus and enabling the study of MT-MMP-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MT-MMP-2 pathway restoration in tumor cells with silenced or reduced MMP15 expression.

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