GIPR CRISPR Activation Plasmid (h): sc-402895-ACT

Human GIPR encodes the glucose-dependent insulinotropic polypeptide receptor, a class B GPCR expressed in pancreatic islets and other metabolically active tissues that couples primarily to Gs to elevate cAMP and activate PKA/CREB-dependent transcription. GIPR signaling modulates glucose-stimulated insulin secretion, β-cell survival, and nutrient-driven anabolic programs, with additional cross-talk to PI3K/AKT and ERK/MAPK pathways that influence cellular growth and stress responses. Altered GIPR expression or signaling has been implicated in metabolic dysregulation, including obesity and type 2 diabetes–related phenotypes, and is also used as a marker of endocrine lineage state. As a result, GIPR is widely studied in hormone receptor biology, islet function, and cAMP-regulated gene networks relevant to metabolic disease mechanisms.

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

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

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