PAF-R CRISPR Activation Plasmid (h): sc-402569-ACT

PTAFR encodes platelet-activating factor receptor (PAF-R), a class A GPCR that binds platelet-activating factor and couples primarily to Gαq/11 and Gαi signaling to regulate phospholipase C activation, intracellular Ca²⁺ flux, and downstream MAPK and NF-κB transcriptional programs. PAF-R activity integrates inflammatory lipid mediator sensing with leukocyte chemotaxis, endothelial activation, and modulation of cytokine and eicosanoid networks, influencing innate immune responses and vascular permeability. Dysregulated PTAFR/PAF-R signaling has been linked to inflammatory and allergic processes, infection-associated immune activation, and tumor microenvironment biology through effects on myeloid cell behavior and barrier function. As a membrane receptor with rapid ligand-driven signaling, it provides a tractable node for dissecting GPCR-mediated inflammation and lipid signaling crosstalk.

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

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

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