GPR50 CRISPR Activation Plasmid (h): sc-402934-ACT

GPR50 encodes an orphan G protein-coupled receptor most closely related to melatonin receptors and is enriched in neuroendocrine tissues, where it is implicated in regulation of circadian and metabolic physiology. Although it does not bind melatonin with high affinity, GPR50 can modulate GPCR signaling through receptor interactions and trafficking, influencing second-messenger pathways that shape neuronal excitability and hormonal responses. Reported associations connect GPR50 variation or altered expression with mood disorders, sleep and seasonal phenotypes, and metabolic traits, supporting its relevance to CNS and energy-balance research. As a membrane receptor-like protein, GPR50 is also used to study GPCR evolution, receptor dimerization, and context-dependent signaling in human cell models.

GPR50 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR50 expression without altering the underlying DNA sequence.

GPR50 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR50 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 GPR50 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR50 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR50 locus and enabling the study of GPR50-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR50 pathway restoration in tumor cells with silenced or reduced GPR50 expression.

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