PDGF-D CRISPR Activation Plasmid (h): sc-402406-ACT

PDGFD encodes platelet-derived growth factor D (PDGF-D), a secreted growth factor that signals primarily through PDGFRβ to regulate mesenchymal cell proliferation, migration, and extracellular matrix remodeling. PDGF-D–PDGFR axis activation engages downstream MAPK/ERK, PI3K/AKT, and STAT pathways that coordinate angiogenesis-related processes and stromal–epithelial crosstalk. Dysregulated PDGFD expression has been linked to altered fibroblast activation, vascular remodeling, and invasive phenotypes in multiple disease-relevant contexts, making it a useful node for studying paracrine signaling in complex microenvironments. In vitro and in vivo models leveraging PDGF-D perturbation are commonly used to interrogate growth factor–driven changes in cell motility, survival, and matrix deposition.

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

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

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