Met CRISPR Activation Plasmid (m): sc-421635-ACT

Mouse Met encodes a receptor tyrosine kinase for hepatocyte growth factor (HGF) that regulates epithelial and endothelial proliferation, survival, and motility through MAPK/ERK, PI3K–AKT, STAT, and Rho-family signaling. MET signaling coordinates invasive growth programs, cytoskeletal remodeling, and cell–cell interactions during embryonic development and tissue repair. Dysregulated MET activity is implicated in oncogenic signaling networks, metastatic dissemination, and altered tumor–stromal communication, and it also intersects with pathways relevant to fibrosis and inflammatory microenvironments. In experimental systems, Met serves as a key node for studying receptor activation dynamics, downstream phosphorylation cascades, and transcriptional programs that control migration and morphogenesis.

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

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

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