The HPCAL1 Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-edited population of A-549 lung adenocarcinoma cells with targeted disruption of the HPCAL1 gene. As a polyclonal knockout pool, this product consists of a bulk-edited cell population without single-cell cloning, providing a heterogeneous model for loss-of-function studies. The disruption of HPCAL1, encoding a neuronal calcium sensor protein, enables the investigation of its roles in GPCR signaling and calcium-dependent regulation.
The A-549 cell line, derived from human lung adenocarcinoma, serves as a widely used model for type II alveolar epithelial cells. Characterized by epithelial morphology and expression of surfactant proteins, A-549 cells are instrumental in studying lung cancer biology, including tumor proliferation, migration, and drug resistance.
HPCAL1 is a calcium-binding protein of the neuronal calcium sensor family. Upon calcium influx, it undergoes conformational changes to interact with GRK2 (G protein-coupled receptor kinase 2), promoting receptor desensitization and internalization. This modulation affects downstream signaling, such as cAMP production downstream of beta-adrenergic receptors. HPCAL1 activity is regulated by calcium signals via GPCR activation or calcium channels, as well as by protein kinase C (PKC). Additionally, it interacts with rhodopsin kinase in phototransduction. Overall, HPCAL1 integrates calcium signaling with GPCR responsiveness, influencing processes like synaptic plasticity and phototransduction.
In A-549 cells, knockout of HPCAL1 allows dissection of its role in calcium-dependent GPCR regulation within a lung adenocarcinoma context. While traditionally studied in neuronal and retinal tissues, HPCAL1 expression in A-549 provides a unique opportunity to explore non-neuronal functions. Aberrant GPCR signaling drives many cancer hallmarks; therefore, loss of HPCAL1 may help elucidate its impact on GRK2-mediated desensitization, tumor cell migration, or drug sensitivity. This model bridges calcium sensor biology with oncogenic pathways, potentially revealing therapeutic targets.
Applications include GPCR internalization assays to measure receptor desensitization, cAMP assays for second messenger quantification, and calcium imaging to monitor intracellular calcium dynamics. Migration assays assess functional consequences in cancer aggressiveness. RNA-seq and RT-qPCR enable transcriptomic profiling and validation of the knockout, while Western blotting confirms protein-level changes in partners like GRK2. These cells are also suitable for drug screens targeting GPCR pathways in lung adenocarcinoma. For more information, contact Ascent Research.
