GPR84 CRISPR Activation Plasmid (h): sc-401953-ACT

Human GPR84 encodes a G protein–coupled receptor that is enriched in myeloid lineages and functions as a sensor for medium-chain fatty acids, linking lipid metabolites to innate immune activation. Upon ligand engagement, GPR84 primarily couples to Gi/o signaling to modulate cAMP levels and downstream pathways that shape chemotaxis, cytokine output, and inflammatory gene programs. It is implicated in macrophage and microglial responses, with reported relevance to chronic inflammatory states and metabolic inflammation. Dysregulated GPR84 signaling has been associated with immune-driven pathology across tissues, supporting its use as a probe for immunometabolic mechanisms.

GPR84 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR84 expression without altering the underlying DNA sequence.

GPR84 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR84 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 GPR84 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR84 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR84 locus and enabling the study of GPR84-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR84 pathway restoration in tumor cells with silenced or reduced GPR84 expression.

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