PGI2 synthase CRISPR Activation Plasmid (h): sc-403180-ACT

Human PTGIS encodes prostacyclin (PGI2) synthase, a cytochrome P450 enzyme that converts prostaglandin H2 to prostacyclin, a lipid mediator that elevates cAMP signaling and modulates platelet function, vascular tone, and inflammatory responses. PTGIS operates within arachidonic acid and eicosanoid biosynthesis pathways, integrating with COX-derived prostanoid production to shape endothelial and smooth muscle cell phenotypes. Altered PTGIS expression or PGI2 balance has been associated with cardiovascular and inflammatory biology, including vascular remodeling and thrombosis-related processes, and is also relevant to tumor microenvironment signaling through prostanoid networks. As such, PTGIS is a useful target for dissecting prostanoid-dependent regulation of endothelial activation, immune cell trafficking, and oxidative stress responses in human cell models.

PGI2 synthase CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTGIS expression without altering the underlying DNA sequence.

PGI2 synthase CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTGIS 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 PTGIS transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PGI2 synthase expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTGIS locus and enabling the study of PGI2 synthase-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PGI2 synthase pathway restoration in tumor cells with silenced or reduced PTGIS expression.

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