GPR174 CRISPR Activation Plasmid (h): sc-413678-ACT

GPR174 (also known as the lysophosphatidylserine receptor) is a G protein-coupled receptor enriched in immune cells and implicated in regulation of lymphocyte activation, cytokine output, and immune homeostasis. Upon ligand engagement it can couple to Gαs-dependent signaling, influencing cAMP production and downstream transcriptional programs that shape T cell and B cell functional states. GPR174 activity intersects with broader GPCR-controlled pathways governing migration, activation thresholds, and inflammatory signaling networks. Dysregulated expression or signaling of GPR174 has been linked to immune-mediated disease biology and provides a mechanistic entry point for studying immunoregulation in human cellular models.

GPR174 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR174 expression without altering the underlying DNA sequence.

GPR174 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR174 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 GPR174 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR174 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR174 locus and enabling the study of GPR174-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR174 pathway restoration in tumor cells with silenced or reduced GPR174 expression.

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