HGF CRISPR Activation Plasmid (h): sc-400433-ACT

Hepatocyte growth factor (HGF) is a secreted pleiotropic cytokine that signals primarily through the MET receptor tyrosine kinase to regulate epithelial–mesenchymal interactions, cell survival, proliferation, motility, and branching morphogenesis. HGF–MET activation engages downstream PI3K–AKT, RAS–MAPK, STAT, and focal adhesion signaling, integrating cues that shape tissue repair and remodeling. Dysregulated HGF expression or pathway hyperactivation is frequently studied in tumor cell invasion, metastasis, angiogenesis, and stromal–tumor crosstalk, as well as in fibrotic and inflammatory microenvironments. As a context-dependent modulator of cell plasticity, HGF is widely used to interrogate paracrine signaling and microenvironment-driven phenotypes in human cell models.

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

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

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