PlGF CRISPR Activation Plasmid (h): sc-402864-ACT

Placenta growth factor (PGF) encodes PlGF, a secreted member of the VEGF family that signals primarily through VEGFR1/FLT1 to regulate angiogenesis, vascular permeability, and endothelial cell survival. PlGF participates in hypoxia-responsive and inflammatory programs that shape vascular remodeling and myeloid cell recruitment, intersecting with VEGF/VEGFR, PI3K–AKT, and MAPK pathway activity. Dysregulated PGF/PlGF expression has been associated with aberrant neovascularization and microenvironmental changes observed in ischemic and inflammatory settings, as well as tumor-associated angiogenic signaling. As a biomarker-linked vascular factor, PlGF is frequently studied in endothelial biology, placental development, and disease-relevant models of pathological vascular growth.

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

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

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