EP1 CRISPR Activation Plasmid (h): sc-402565-ACT

PTGER1 encodes the human prostaglandin E2 receptor EP1, a G protein–coupled receptor that preferentially couples to Gq/11 to elevate intracellular Ca2+ and activate PKC-dependent signaling. EP1 integrates prostanoid inputs downstream of cyclooxygenase pathways to regulate smooth muscle tone, epithelial ion transport, and neuroinflammatory responses, with additional cross-talk into MAPK and calcium-dependent transcriptional programs. Altered PTGER1/EP1 signaling has been studied in inflammation-driven tissue remodeling and pain sensitization, and it has been implicated in context-dependent regulation of cell proliferation and migration. These features make EP1 a useful node for probing PGE2-responsive signaling networks and downstream gene expression changes in human cell models.

EP1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTGER1 expression without altering the underlying DNA sequence.

EP1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTGER1 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 PTGER1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous EP1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTGER1 locus and enabling the study of EP1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of EP1 pathway restoration in tumor cells with silenced or reduced PTGER1 expression.

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