The CCDC82 Knockout NCI-H1975 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population generated by disruption of the CCDC82 gene in the human NCI-H1975 lung adenocarcinoma cell line. This polyclonal knockout model provides a heterogeneous pool of edited cells, enabling the study of CCDC82 loss-of-function effects without clonal selection bias, and is suitable for functional genomics and drug discovery applications requiring robust gene knockout in a cancer-relevant background.
The NCI-H1975 cell line is a widely used model of non-small cell lung cancer (NSCLC) derived from a lung adenocarcinoma patient. These epithelial cells harbor the EGFR T790M mutation, which confers resistance to first-generation tyrosine kinase inhibitors (TKIs) such as gefitinib and erlotinib. Consequently, NCI-H1975 cells are a standard platform for investigating mechanisms of acquired drug resistance and for evaluating next-generation EGFR inhibitors. The integration of CCDC82 knockout into this background allows dissection of gene function specifically within the context of EGFR-mutant lung cancer.
CCDC82 encodes a coiled-coil domain-containing protein with predicted functions in centrosome biology and cell cycle control. Although its exact regulatory mechanisms remain poorly defined, available evidence suggests that CCDC82 localizes to the centrosome and participates in microtubule organization. The protein is proposed to interact with centrosomal scaffold components such as CEP152 and CEP63, as well as centriolar satellite proteins like PCM1 and CEP131. Downstream, CCDC82 may influence cell cycle progression and apoptosis, potentially through interactions with cyclin-dependent kinases. Upstream regulation is thought to involve cell cycle transcription factors, consistent with its role in proliferation control.
In NCI-H1975 cells, knockout of CCDC82 is expected to compromise centrosome integrity and function, leading to aberrant cell cycle progression, mitotic defects, and altered apoptotic responses. This model is particularly valuable for exploring the contribution of centrosomal defects to lung adenocarcinoma tumorigenesis and for assessing how centrosome dysfunction intersects with EGFR signaling and drug resistance. By providing a loss-of-function system in an EGFR T790M-positive background, the polyclonal knockout cells facilitate the identification of synthetic lethal interactions and the testing of therapeutic strategies that target cell cycle vulnerabilities in NSCLC.
These polyclonal knockout cells are suitable for a broad range of experimental applications. Researchers can employ Western blotting and RNA-seq transcriptomic profiling to validate knockout efficiency and assess global gene expression changes. Cell viability assays (MTT or CellTiter-Glo) and apoptosis assays (Annexin V/caspase activation) enable quantification of proliferative and survival phenotypes. Cell cycle analysis by flow cytometry can reveal mitotic delays or G1/S arrest, while immunofluorescence microscopy allows visualization of centrosome and microtubule anomalies. Co-immunoprecipitation studies help elucidate CCDC82 protein interaction networks, and drug sensitivity assays with EGFR inhibitors can probe the impact of CCDC82 loss on therapeutic response. For additional information and technical support, please contact Ascent Research.
