The CCK Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-mediated polyclonal knockout population of HAP1 cells, engineered to disrupt the CCK gene and eliminate cholecystokinin (CCK) peptide expression. This product delivers a heterogeneous cell pool suitable for loss-of-function studies, enabling researchers to investigate CCK-dependent biology without the confounding effects of endogenous hormone production. The polyclonal nature ensures a broad representation of edited cells while circumventing the need for monoclonal isolation and expansion.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and exhibit a near-haploid karyotype with adherent fibroblastoid morphology. The haploid genome facilitates efficient gene editing, as disruption of a single allele yields functional knockout. This characteristic, combined with robust expansion, makes HAP1 an ideal host for genetic perturbation studies, including high-throughput screens and detailed signaling analyses.
The CCK gene product is a multifunctional peptide hormone and neurotransmitter acting via two GPCRs, CCKAR and CCKBR. Ligand binding triggers G??q-mediated PLC?? activation, producing IP3 and DAG, which mobilize intracellular Ca2? and activate PKC. This primary cascade feeds into the RAF-MEK-ERK1/2 and PI3K-AKT pathways, culminating in CREB phosphorylation. Physiologically, CCK release is stimulated by dietary fatty acids, amino acids, and vagal input, while processing by prohormone convertases (PC1/3, PC2) generates active forms. The system closely interacts with gastrin and proteases.
In the HAP1 background, CCK knockout creates a clean cellular model to dissect receptor pharmacology and signaling without endogenous CCK interference. Although myeloid-derived, these cells support exogenous receptor expression and ligand-induced responses, enabling precise measurement of second messengers and kinase activation. Researchers can employ calcium mobilization assays, phospho-ERK western blotting, and luciferase reporter systems to evaluate CCK analog efficacy or screen compound libraries targeting CCKAR/CCKBR.
Key applications include GPCR deorphanization, hormone function studies, satiety regulation research, and gastrointestinal cancer modeling. Assays such as RT-qPCR for CCK transcript, ELISA for secreted peptide, flow cytometry for receptor surface expression, and drug sensitivity testing with gastrin/CCK are fully compatible. These polyclonal knockout cells serve as a versatile platform for exploring MAPK/ERK and PI3K-AKT signaling in contexts like obesity, anxiety disorders, and pancreatitis. For technical inquiries, please contact Ascent Research.
