GLP-2R CRISPR Activation Plasmid (h): sc-403966-ACT

GLP2R encodes the glucagon-like peptide-2 receptor (GLP-2R), a class B GPCR predominantly expressed in intestinal and enteric tissues where it senses the incretin hormone GLP-2. Upon ligand engagement, GLP-2R couples primarily to Gs to elevate cAMP and activate PKA- and CREB-associated transcriptional programs that regulate epithelial growth, barrier integrity, nutrient absorption, and mucosal repair. Downstream signaling intersects with pathways controlling cell survival and proliferation, including MAPK and PI3K-linked processes, shaping intestinal homeostasis and neuroendocrine communication. Dysregulated GLP2R signaling has been studied in contexts of gastrointestinal inflammation, malabsorption, and altered epithelial remodeling, supporting mechanistic research into gut barrier function and metabolic physiology.

GLP-2R CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GLP2R expression without altering the underlying DNA sequence.

GLP-2R CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GLP2R 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 GLP2R transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GLP-2R expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GLP2R locus and enabling the study of GLP-2R-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GLP-2R pathway restoration in tumor cells with silenced or reduced GLP2R expression.

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