GPR20 CRISPR Activation Plasmid (h): sc-407471-ACT

GPR20 encodes an orphan class A G protein-coupled receptor predominantly linked to neuronal and neuroendocrine biology, where it is thought to shape receptor-mediated signaling that influences cyclic AMP dynamics, MAPK activity, and downstream transcriptional programs. As a membrane GPCR, GPR20 can modulate cell–cell communication and stimulus-responsive signaling networks that coordinate differentiation, excitability, and secretory processes in relevant cellular contexts. Altered GPCR signaling is broadly implicated in neuropsychiatric and metabolic phenotypes, making GPR20 a useful target for dissecting pathway connectivity and receptor-driven regulatory states. Human GPR20 is therefore of interest for mechanistic studies of GPCR-associated signal transduction and for mapping gene network effects in disease-relevant model systems.

GPR20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR20 expression without altering the underlying DNA sequence.

GPR20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR20 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 GPR20 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR20 locus and enabling the study of GPR20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR20 pathway restoration in tumor cells with silenced or reduced GPR20 expression.

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