FOXD1 CRISPR Activation Plasmid (h): sc-402521-ACT

FOXD1 (forkhead box D1) is a forkhead family transcription factor that regulates cell fate specification, proliferation, and differentiation programs during embryonic development, with prominent roles in kidney morphogenesis and stromal lineage patterning. By binding forkhead DNA motifs, FOXD1 coordinates transcriptional networks that intersect with developmental signaling pathways such as WNT, TGF-β, and growth factor–driven MAPK/PI3K signaling to shape tissue architecture. Dysregulated FOXD1 expression has been associated with altered epithelial–stromal interactions, extracellular matrix remodeling, and invasive cellular phenotypes in multiple cancer contexts, making it a useful node for studying transcriptional control of tumor microenvironmental programs. In addition, FOXD1-linked gene regulatory circuits are relevant to investigating developmental disorders and lineage-specific transcriptional states in human cells.

FOXD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXD1 expression without altering the underlying DNA sequence.

FOXD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXD1 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 FOXD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous FOXD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXD1 locus and enabling the study of FOXD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of FOXD1 pathway restoration in tumor cells with silenced or reduced FOXD1 expression.

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