FOXC1 CRISPR Activation Plasmid (m): sc-421637-ACT

Foxc1 encodes the forkhead box transcription factor FOXC1, a sequence-specific DNA-binding regulator that controls developmental patterning, cell fate decisions, and differentiation programs. In mouse systems, FOXC1 integrates signals across transcriptional networks involved in mesenchymal–epithelial interactions, extracellular matrix remodeling, and morphogen-driven pathways such as TGF-β/BMP and WNT to shape organogenesis and tissue homeostasis. Altered FOXC1 dosage or regulatory activity is linked to congenital anterior segment and craniofacial phenotypes and has been associated with dysregulated cell identity and invasive gene expression signatures in cancer biology. These properties make FOXC1 a useful node for dissecting gene regulatory circuits governing development, stem-like states, and microenvironmental responses.

FOXC1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Foxc1 expression without altering the underlying DNA sequence.

FOXC1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Foxc1 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 Foxc1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXC1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Foxc1 locus and enabling the study of FOXC1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXC1 pathway restoration in tumor cells with silenced or reduced Foxc1 expression.

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