Glucagon Receptor CRISPR Activation Plasmid (h): sc-402018-ACT

GCGR encodes the human glucagon receptor, a class B G protein–coupled receptor that binds glucagon and couples primarily to Gαs to stimulate adenylate cyclase, elevate cAMP, and activate PKA-dependent transcriptional programs. This signaling axis regulates hepatic glucose production, glycogenolysis, and lipid metabolism, and intersects with CREB-mediated gene expression, metabolic stress responses, and endocrine control of energy homeostasis. Altered GCGR expression or signaling has been implicated in dysregulated glucose and lipid handling and is frequently studied in the context of metabolic disease biology. As a membrane receptor with well-defined downstream readouts, GCGR serves as a tractable node for dissecting hormone-driven pathways in hepatocytes and other relevant cell types.

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

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

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