glypican-1 CRISPR Activation Plasmid (h): sc-402002-ACT

Human GPC1 encodes glypican-1, a glycosylphosphatidylinositol-anchored heparan sulfate proteoglycan that localizes to the cell surface and binds diverse growth factors, morphogens, and extracellular matrix components. By modulating ligand availability and receptor engagement, glypican-1 influences signaling outputs in pathways such as FGF, VEGF, Wnt, Hedgehog, and TGF-β, with downstream effects on proliferation, adhesion, migration, and angiogenic programs. Altered GPC1 expression and glypican-1 shedding have been associated with changes in tumor microenvironment communication, invasive phenotypes, and biomarker studies in multiple cancer contexts. Beyond oncology, glypican-1–dependent regulation of heparan sulfate interactions is relevant to neurodevelopmental processes and cellular responses to injury and inflammation.

glypican-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPC1 expression without altering the underlying DNA sequence.

glypican-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPC1 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 GPC1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous glypican-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPC1 locus and enabling the study of glypican-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of glypican-1 pathway restoration in tumor cells with silenced or reduced GPC1 expression.

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