DP2 CRISPR Activation Plasmid (h): sc-404990-ACT

Human PTGDR2 encodes DP2 (CRTH2), a G protein–coupled receptor for prostaglandin D2 that is enriched in type 2 immune cell populations including eosinophils, basophils, and Th2 lymphocytes. DP2 signaling couples primarily to Gi-dependent pathways to regulate chemotaxis, calcium mobilization, and cytokine programs that shape allergic inflammation and mucosal immune responses. Through modulation of leukocyte trafficking and effector function, PTGDR2 activity is frequently investigated in the context of asthma, allergic rhinitis, atopic dermatitis, and eosinophilic inflammatory states. As a cell-surface receptor integrating lipid mediator cues, DP2 provides a tractable node for studying GPCR signal transduction and immune cell migration circuits.

DP2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTGDR2 expression without altering the underlying DNA sequence.

DP2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTGDR2 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 PTGDR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DP2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTGDR2 locus and enabling the study of DP2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DP2 pathway restoration in tumor cells with silenced or reduced PTGDR2 expression.

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