TIMP-1 CRISPR Activation Plasmid (h): sc-400408-ACT

TIMP1 encodes tissue inhibitor of metalloproteinases-1 (TIMP-1), a secreted glycoprotein that binds and inhibits multiple matrix metalloproteinases to regulate extracellular matrix turnover and pericellular proteolysis. By constraining MMP activity, TIMP-1 influences cell migration, adhesion, and tissue remodeling, and intersects with signaling networks that couple matrix dynamics to cellular responses, including inflammatory and fibrotic processes. Dysregulated TIMP1 expression is frequently studied in contexts of aberrant matrix remodeling such as fibrosis, chronic inflammation, and tumor microenvironment biology, where altered ECM homeostasis can affect invasion and metastasis-associated phenotypes. As an X-linked gene with broad expression, TIMP1 also serves as a useful marker for investigating protease–antiprotease balance across diverse human cell types.

TIMP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TIMP1 expression without altering the underlying DNA sequence.

TIMP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TIMP1 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 TIMP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TIMP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TIMP1 locus and enabling the study of TIMP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TIMP-1 pathway restoration in tumor cells with silenced or reduced TIMP1 expression.

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