This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from A-549 cells, featuring targeted disruption of the CCDC50 gene. As a polyclonal pool, the knockout cells reflect a heterogeneous collection of edited alleles, providing a robust loss-of-function model for studying CCDC50-dependent processes without the selection biases associated with single-cell clones. The use of CRISPR/Cas9 technology ensures efficient gene disruption, enabling researchers to interrogate the functional role of CCDC50 in relevant cellular contexts. This population is suitable for a wide range of downstream applications, including signaling pathway analysis, protein turnover assays, and functional genomics studies in lung adenocarcinoma models.
The parental A-549 cell line is a human lung adenocarcinoma epithelial line originally derived from a 58-year-old Caucasian male. A-549 cells are widely utilized as an in vitro model system for non-small cell lung cancer (NSCLC) research, drug screening, and mechanistic studies of oncogenic signaling. These adherent cells express wild-type EGFR and retain key characteristics of the alveolar epithelium, making them particularly valuable for investigating growth factor receptor biology. Their well-characterized response to EGF stimulation and established use in assays measuring cell proliferation, migration, and apoptosis provide a consistent and physiologically relevant background for CCDC50 knockout experiments.
CCDC50 encodes coiled-coil domain-containing protein 50, a cytoplasmic adaptor that functions as a negative regulator of epidermal growth factor receptor (EGFR) signaling. Mechanistically, CCDC50 bridges ligand-activated EGFR and the CBL ubiquitin ligase, facilitating the ubiquitination of EGFR and subsequent sorting into the endosomal-lysosomal degradation pathway via the ESCRT machinery. This activity directly controls EGFR protein stability and attenuates downstream signaling cascades, including MAPK/ERK and PI3K/AKT phosphorylation. In addition, CCDC50 modulates NF-??B transcriptional activity through interactions with TAB2 and TAK1, linking EGFR trafficking to inflammatory signaling. Key interacting partners include EGFR, CBL, GRB2, TAB2, and TAK1, underscoring its role at the intersection of receptor endocytosis, ubiquitin-mediated proteolysis, and intracellular signal transduction.
In the A-549 lung adenocarcinoma cell background, CCDC50 serves as a critical checkpoint for EGFR signal duration and intensity. Loss of CCDC50 disrupts the normal ligand-induced degradation of EGFR, leading to sustained receptor expression, prolonged phosphorylation of ERK and AKT, and enhanced NF-??B activation. This dysregulation mimics oncogenic signaling patterns frequently observed in NSCLC, where aberrant EGFR activity drives tumor growth, survival, and therapy resistance. Consequently, this knockout model recapitulates a state of heightened proliferative and survival signaling, providing a powerful tool for dissecting the molecular mechanisms that underlie EGFR-driven lung cancer progression and for identifying vulnerabilities that may be exploitable therapeutically.
Researchers can employ this polyclonal knockout population in a variety of experimental settings. It is ideal for EGF pulse-chase assays to quantify EGFR degradation kinetics, ubiquitination assays to probe CBL-mediated receptor modification, and immunoblotting to assess changes in phospho-EGFR, phospho-ERK, phospho-AKT, and NF-??B pathway components. Functional studies may include colony formation assays, MTT-based proliferation measurements, and migration/invasion assays to evaluate cellular phenotypes associated with enhanced EGFR output. This model also supports drug screening for modulators of EGFR trafficking and for testing therapeutic combinations in the context of NSCLC. For additional information or technical support, please contact Ascent Research.
