GPR105 CRISPR Activation Plasmid (h): sc-405954-ACT

P2RY14 encodes the human purinergic receptor GPR105, a GPCR that responds to extracellular UDP-sugars such as UDP-glucose and couples to Gi/o signaling to modulate intracellular cAMP and downstream inflammatory programs. GPR105 activity influences leukocyte chemotaxis and cytokine production, linking nucleotide-sugar sensing to innate immune regulation in myeloid and epithelial compartments. This receptor intersects with chemokine and purinergic signaling networks that shape tissue inflammation and barrier responses. Altered P2RY14/GPR105 signaling has been associated with immune dysregulation and inflammation-related pathophysiology, making it relevant for mechanistic studies of microenvironment-driven disease processes.

GPR105 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous P2RY14 expression without altering the underlying DNA sequence.

GPR105 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the P2RY14 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 P2RY14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR105 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native P2RY14 locus and enabling the study of GPR105-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR105 pathway restoration in tumor cells with silenced or reduced P2RY14 expression.

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