CCK-AR CRISPR Activation Plasmid (h): sc-402419-ACT

Human CCKAR encodes the cholecystokinin A receptor (CCK-AR), a class A GPCR activated by cholecystokinin peptides that regulates digestion-associated signaling and nutrient-driven satiety cues. Upon ligand binding, CCK-AR engages G protein pathways that modulate intracellular calcium and downstream kinase cascades, coordinating secretory responses and smooth muscle contractility in gastrointestinal tissues. CCKAR activity intersects with neuroendocrine communication between gut and brain and influences cellular programs linked to metabolism and energy balance. Dysregulated CCKAR signaling has been investigated in the context of metabolic phenotypes and gastrointestinal functional alterations, supporting its relevance for mechanistic studies of receptor-mediated signal transduction.

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

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

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