GPRC6A CRISPR Activation Plasmid (h): sc-403492-ACT

GPRC6A encodes a class C G protein-coupled receptor implicated in nutrient sensing and endocrine-metabolic crosstalk, responding to extracellular cues such as basic amino acids and divalent cations. Through coupling to heterotrimeric G proteins, GPRC6A can modulate second messenger signaling and downstream kinase pathways that influence cellular metabolism, hormone secretion, and inflammatory tone. Its activity has been studied in contexts including glucose homeostasis, energy balance, and osteoendocrine signaling, linking receptor-dependent signaling to metabolic and skeletal biology. Altered GPRC6A expression or signaling has been associated in the literature with complex disease phenotypes relevant to obesity, type 2 diabetes, and cardiometabolic traits, supporting its use in mechanistic pathway studies.

GPRC6A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPRC6A expression without altering the underlying DNA sequence.

GPRC6A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPRC6A 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 GPRC6A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPRC6A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPRC6A locus and enabling the study of GPRC6A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPRC6A pathway restoration in tumor cells with silenced or reduced GPRC6A expression.

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