proglucagon CRISPR Activation Plasmid (h): sc-416721-ACT

Human GCG encodes proglucagon, a prohormone processed in a tissue-specific manner to yield glucagon in pancreatic α cells and incretin peptides such as GLP‑1 and GLP‑2 in intestinal L cells and select neurons. These peptides coordinate systemic energy balance by regulating hepatic glucose output, insulin secretion, appetite signaling, and intestinal growth, linking GCG to endocrine and metabolic homeostasis. Proglucagon-derived hormones signal primarily through GPCR pathways that engage cAMP/PKA and CREB-dependent transcriptional programs, integrating nutrient sensing with hormonal responses. Dysregulated proglucagon processing or signaling is associated with impaired glucose regulation and broader metabolic disease phenotypes, making GCG a key node for studying islet biology and gut–pancreas endocrine axes.

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

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

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