GIP CRISPR Activation Plasmid (h): sc-402545-ACT

Human GIP encodes glucose-dependent insulinotropic polypeptide, an incretin hormone produced by enteroendocrine K cells that amplifies glucose-stimulated insulin secretion and influences glucagon release, gastric motility, and nutrient handling. GIP signaling through the GIP receptor engages cAMP/PKA and downstream transcriptional programs that coordinate pancreatic islet function and systemic energy balance. Dysregulation of GIP axis activity is implicated in metabolic phenotypes, including impaired incretin responses, altered adipose lipid metabolism, and obesity-associated insulin resistance, making GIP a useful node for interrogating enteroinsular communication. Experimental modulation of GIP expression supports mechanistic studies of islet hormone cross-talk, nutrient-sensing pathways, and metabolic stress responses in relevant cellular models.

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

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

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