PGDH CRISPR Activation Plasmid (h): sc-402622-ACT

Human HPGD encodes 15-hydroxyprostaglandin dehydrogenase (PGDH), the rate-limiting enzyme responsible for NAD+-dependent oxidation and inactivation of bioactive prostaglandins such as PGE2. By controlling prostanoid turnover, PGDH modulates prostaglandin/COX signaling outputs that influence inflammation, epithelial homeostasis, angiogenic cues, and tissue remodeling. Altered HPGD activity shifts local eicosanoid balance and is frequently studied in contexts where dysregulated prostaglandin metabolism contributes to aberrant immune signaling and proliferative phenotypes. As a gatekeeper of prostaglandin catabolism, HPGD is a useful node for dissecting crosstalk between inflammatory mediators, metabolic state, and transcriptional programs.

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

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

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