FOXF1 CRISPR Activation Plasmid (h): sc-403521-ACT

FOXF1 (forkhead box F1) encodes a winged-helix transcription factor that regulates mesenchymal differentiation and organogenesis, with prominent roles in pulmonary and vascular development. In the nucleus, FOXF1 coordinates gene programs controlling cell migration, extracellular matrix remodeling, and smooth muscle/pericyte-associated transcriptional networks, integrating developmental signaling inputs such as Hedgehog and TGF-β pathway activity. Altered FOXF1 dosage or regulatory disruption is linked to congenital lung and vascular disorders, and aberrant expression has been studied in contexts of tissue remodeling and cancer-associated stromal biology. These features make FOXF1 a useful node for dissecting transcriptional circuitry governing mesenchymal cell state transitions and epithelial–mesenchymal interactions.

FOXF1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXF1 expression without altering the underlying DNA sequence.

FOXF1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXF1 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 FOXF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXF1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXF1 locus and enabling the study of FOXF1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXF1 pathway restoration in tumor cells with silenced or reduced FOXF1 expression.

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