PGF2αR CRISPR Activation Plasmid (h): sc-402974-ACT

PTGFR encodes the prostaglandin F2α receptor (PGF2αR), a rhodopsin-like GPCR that binds PGF2α to initiate Gq/11-dependent signaling, elevating intracellular calcium and activating phospholipase C, PKC, and downstream MAPK pathways. Through these cascades, PGF2αR regulates smooth muscle contractility, vascular tone, and context-dependent inflammatory responses, linking prostanoid biology to tissue remodeling programs. Altered PTGFR signaling has been studied in reproductive physiology, ocular pressure regulation, and cancer-associated microenvironmental cues, where prostaglandin-driven signaling can influence proliferation, migration, and extracellular matrix dynamics. As a membrane receptor integrating eicosanoid signals, PGF2αR serves as a tractable node for interrogating GPCR-mediated transcriptional and phenotypic outputs.

PGF2αR CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTGFR expression without altering the underlying DNA sequence.

PGF2αR CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTGFR 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 PTGFR transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PGF2αR expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTGFR locus and enabling the study of PGF2αR-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PGF2αR pathway restoration in tumor cells with silenced or reduced PTGFR expression.

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