GPRC5B CRISPR Activation Plasmid (h): sc-409271-ACT

GPRC5B (G protein-coupled receptor class C group 5 member B) is an orphan GPCR-like transmembrane protein implicated in modulation of receptor signaling at the plasma membrane and endomembrane compartments. It has been linked to regulation of MAPK/ERK and PI3K–AKT-associated processes that shape proliferation, differentiation, and cellular stress responses, with context-dependent effects on inflammatory signaling and metabolic homeostasis. Expression changes in GPRC5B have been reported across multiple tumor types and in disorders featuring dysregulated immune or metabolic pathways, supporting its utility as a node for studying signal integration. As a result, GPRC5B is frequently examined in models of oncogenic signaling, adipocyte and immune cell biology, and receptor cross-talk influencing cell state transitions.

GPRC5B CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPRC5B expression without altering the underlying DNA sequence.

GPRC5B CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPRC5B 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 GPRC5B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPRC5B expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPRC5B locus and enabling the study of GPRC5B-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPRC5B pathway restoration in tumor cells with silenced or reduced GPRC5B expression.

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