FOXC2 CRISPR Activation Plasmid (m): sc-420370-ACT

Forkhead box protein C2 (FOXC2), encoded by the mouse Foxc2 gene, is a winged-helix transcription factor that orchestrates developmental and tissue homeostasis programs through context-dependent regulation of gene expression. FOXC2 is implicated in vascular and lymphatic development, mesenchymal cell fate decisions, and epithelial–mesenchymal transition-like processes, intersecting with pathways such as TGF-β, Notch, and Wnt signaling that shape morphogenesis and remodeling. Altered FOXC2 activity is associated with defects in lymphatic and cardiovascular biology and has been linked to dysregulated cell migration and invasiveness in disease-associated models. These properties make Foxc2 a useful node for studying transcriptional networks controlling differentiation, barrier function, and tissue remodeling.

FOXC2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Foxc2 expression without altering the underlying DNA sequence.

FOXC2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Foxc2 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 Foxc2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXC2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Foxc2 locus and enabling the study of FOXC2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXC2 pathway restoration in tumor cells with silenced or reduced Foxc2 expression.

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