Mts1 CRISPR Activation Plasmid (h): sc-401774-ACT

S100A4 (Mts1) is a calcium-binding S100 family protein that functions as a regulator of cytoskeletal dynamics, cell motility, and extracellular matrix remodeling. By interacting with nonmuscle myosin II and other cytoskeleton-associated partners, S100A4 influences actin organization and pathways linked to epithelial–mesenchymal transition, invasion, and inflammatory cell trafficking. Altered S100A4 expression is frequently associated with fibrotic remodeling, chronic inflammation, and metastatic progression in multiple tumor types, making it a useful marker and mechanistic node for studying cellular plasticity. In human model systems, S100A4 supports research into migration, adhesion turnover, and microenvironmental signaling that shapes tissue remodeling and disease phenotypes.

Mts1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous S100A4 expression without altering the underlying DNA sequence.

Mts1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the S100A4 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 S100A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Mts1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native S100A4 locus and enabling the study of Mts1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Mts1 pathway restoration in tumor cells with silenced or reduced S100A4 expression.

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