PGD synthase CRISPR Activation Plasmid (h): sc-405437-ACT

Human HPGDS encodes hematopoietic prostaglandin D synthase (PGD synthase), a glutathione-dependent enzyme that converts prostaglandin H2 to prostaglandin D2 (PGD2). PGD2 is a bioactive lipid mediator that signals through DP1/DP2 receptors and is further metabolized to cyclopentenone prostaglandins, linking HPGDS activity to eicosanoid metabolism, inflammatory signaling, and redox-sensitive cellular programs. HPGDS expression is prominent in immune and myeloid lineages and has been studied in the context of allergic inflammation, airway and skin immune responses, and neuroinflammatory processes. Altered PGD2 pathway flux can influence leukocyte recruitment, cytokine networks, and tissue remodeling, making HPGDS a useful node for mechanistic studies of inflammation-associated disease phenotypes.

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

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

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