GPR17 CRISPR Activation Plasmid (h): sc-402977-ACT

GPR17 encodes a G protein-coupled receptor implicated in purinergic and cysteinyl leukotriene signaling, integrating extracellular nucleotide and lipid mediator cues into intracellular second-messenger pathways. In the central nervous system, GPR17 is linked to oligodendrocyte lineage progression and myelination programs, influencing cellular responses to injury-associated signals and inflammatory microenvironments. Its activity intersects with GPCR-regulated networks that modulate cyclic nucleotide signaling, MAPK cascades, and chemotactic or stress-response processes depending on cellular context. Dysregulated GPR17 expression or signaling has been investigated in neuroinflammatory and demyelinating conditions, making it a relevant target for mechanistic studies of glial biology and damage-associated signaling.

GPR17 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR17 expression without altering the underlying DNA sequence.

GPR17 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR17 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 GPR17 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR17 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR17 locus and enabling the study of GPR17-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR17 pathway restoration in tumor cells with silenced or reduced GPR17 expression.

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