GLP-1R CRISPR Activation Plasmid (h): sc-400847-ACT

GLP1R encodes the glucagon-like peptide-1 receptor (GLP-1R), a class B GPCR that couples primarily to Gs to stimulate adenylyl cyclase, elevate cAMP, and activate PKA and EPAC signaling. In pancreatic β cells, GLP-1R signaling potentiates glucose-stimulated insulin secretion and modulates gene programs linked to nutrient sensing and secretory capacity, while in extra-pancreatic tissues it influences neuronal and gastrointestinal regulatory circuits. Downstream pathway crosstalk can involve CREB-dependent transcription, PI3K–AKT signaling, and calcium-dependent processes that shape cellular excitability and secretion. Altered GLP1R expression or signaling has been studied in the context of metabolic dysregulation, including diabetes and obesity-associated phenotypes, as well as β-cell stress responses.

GLP-1R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GLP1R expression without altering the underlying DNA sequence.

GLP-1R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GLP1R 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 GLP1R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GLP-1R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GLP1R locus and enabling the study of GLP-1R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GLP-1R pathway restoration in tumor cells with silenced or reduced GLP1R expression.

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