glypican-4 CRISPR Activation Plasmid (m): sc-420640-ACT

Mouse Gpc4 encodes glypican-4, a glycosylphosphatidylinositol-anchored heparan sulfate proteoglycan that localizes to the cell surface and extracellular matrix, where it modulates ligand availability and receptor engagement. Glypican-4 shapes morphogen and growth factor gradients and can influence signaling pathways such as Wnt/β-catenin, Hedgehog, FGF, and BMP through heparan sulfate–dependent interactions. Through these functions, Gpc4 contributes to developmental patterning, tissue remodeling, and cell fate decisions, and altered glypican-mediated signaling has been associated with dysregulated proliferation and differentiation in disease-relevant contexts. In mouse models, Gpc4 is also studied in relation to metabolic and inflammatory phenotypes, including adipose biology and insulin-responsive signaling networks.

glypican-4 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Gpc4 expression without altering the underlying DNA sequence.

glypican-4 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Gpc4 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 Gpc4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous glypican-4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Gpc4 locus and enabling the study of glypican-4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of glypican-4 pathway restoration in tumor cells with silenced or reduced Gpc4 expression.

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