GPR119 CRISPR Activation Plasmid (h): sc-403135-ACT

GPR119 encodes a class A G protein-coupled receptor enriched in pancreatic islet cells and intestinal enteroendocrine populations, where it functions as a lipid-sensing receptor for endogenous fatty acid derivatives. Upon activation, GPR119 primarily couples to Gs to increase intracellular cAMP, engaging PKA- and EPAC-linked signaling that modulates stimulus–secretion coupling and cellular metabolic responses. This receptor is studied in the context of nutrient-induced signaling, incretin-related endocrine communication, and cAMP-dependent transcriptional programs. Dysregulated GPCR and cAMP signaling involving GPR119 has been explored in metabolic disease biology, including pathways relevant to glucose homeostasis and energy balance.

GPR119 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GPR119 expression without altering the underlying DNA sequence.

GPR119 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GPR119 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 GPR119 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GPR119 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GPR119 locus and enabling the study of GPR119-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GPR119 pathway restoration in tumor cells with silenced or reduced GPR119 expression.

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