CCK-BR CRISPR Activation Plasmid (h): sc-401867-ACT

Human CCKBR encodes the cholecystokinin B receptor (CCK-BR), a class A GPCR activated by gastrin and cholecystokinin peptides that couples primarily to Gq/11 to stimulate phospholipase C, IP3/DAG signaling, intracellular Ca2+ flux, and PKC-dependent transcriptional programs. Downstream effects include modulation of MAPK/ERK signaling, cytoskeletal dynamics, and secretory and proliferative responses in gastrointestinal and neuronal contexts. CCKBR activity integrates neuroendocrine cues with epithelial homeostasis and excitability, and dysregulated receptor signaling or expression has been studied in relation to aberrant growth signaling and inflammation-associated remodeling. As a membrane receptor with defined ligand–receptor pharmacology, CCK-BR is also used to interrogate GPCR desensitization, trafficking, and second-messenger pathway bias.

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

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

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