MTSS1 CRISPR Activation Plasmid (h): sc-404758-ACT

Human MTSS1 (metastasis suppressor 1) encodes an I-BAR domain–containing adaptor that links phosphoinositide-rich membranes to actin regulatory networks, shaping membrane curvature and coordinating cytoskeletal remodeling. MTSS1 participates in processes including lamellipodia and filopodia dynamics, endocytosis, and cell–cell adhesion through interactions with actin-associated factors and small GTPase–dependent signaling. By modulating cell motility and invasive behavior, MTSS1 is frequently studied in the context of epithelial differentiation, tumor progression, and metastasis-associated phenotypes. Altered MTSS1 expression has also been connected to changes in developmental programs and stress-responsive transcriptional states relevant to cancer biology and neurobiology research models.

MTSS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MTSS1 expression without altering the underlying DNA sequence.

MTSS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MTSS1 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 MTSS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MTSS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MTSS1 locus and enabling the study of MTSS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MTSS1 pathway restoration in tumor cells with silenced or reduced MTSS1 expression.

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