The CCKAR Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to eliminate functional expression of the human CCKAR gene. This heterogenous pool contains HAP1 cells bearing diverse Cas9-induced disruptions across the CCKAR locus, providing a robust loss-of-function model that avoids clonal selection artifacts. Intended for advanced biomedical research, this product enables rigorous interrogation of cholecystokinin receptor signaling in a genetically simplified background.
HAP1 cells are a near-haploid, adherent human cell line derived from the KBM-7 chronic myeloid leukemia (CML) line. Their haploid genetic architecture reduces the complexity of gene redundancy, ensuring that knockout phenotypes reflect direct gene disruption. As a CML model, HAP1 retains oncogenic signaling features relevant to hematological malignancy studies, while its suitability for high-throughput screening and CRISPR genome editing makes it a preferred platform for GPCR signaling research.
The CCKAR gene encodes a Gq-coupled GPCR that mediates the physiological effects of cholecystokinin (CCK) and, with lower potency, gastrin. Ligand-bound CCKAR activates G??q/11, which stimulates phospholipase C-?? (PLC??) to produce the second messengers IP3 and DAG. IP3 triggers Ca2+ mobilization from the endoplasmic reticulum, while DAG activates protein kinase C (PKC). These events propagate signals through ERK1/2 and CaMK, culminating in the transcriptional induction of immediate-early genes such as FOS and JUN. CCKAR also engages G??s and G??i/o proteins and recruits ??-arrestin-1/2 to modulate receptor desensitization and endocytosis. This canonical pathway regulates gallbladder contraction, pancreatic exocrine secretion, and central satiety circuits.
In the HAP1 background, CCKAR disruption provides a clean, near-haploid system to dissect CCK-dependent signaling events without interference from a second functional allele. This model is particularly powerful for structure?Cfunction analyses of receptor coupling, biased agonism, and trafficking. Moreover, the CML-derived setting offers a unique avenue to probe potential CCKAR functions in hematopoietic cell signaling and leukemogenesis. The polyclonal nature of the knockout population preserves biological variability while ensuring robust loss-of-function across the cell pool, making it ideal for population-based assays and screens.
Key applications include CCK-induced calcium flux assays, phospho-ERK western blotting, and quantitative PCR for FOS/JUN expression to assess downstream signaling competency. The cells are equally suited for receptor binding, internalization, and high-throughput GPCR signaling screens. As a validated tool, they support drug target validation in obesity, gallstone disease, pancreatitis, and eating disorders. For additional technical information or ordering assistance, please contact Ascent Research.
