The CCDC50 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1299 non-small cell lung cancer (NSCLC) cell line, engineered for loss-of-function studies of the human CCDC50 gene. This product comprises a heterogeneous pool of gene-edited cells, enabling robust population-level analysis without the constraints of single-cell cloning. CRISPR/Cas9-mediated targeted disruption of CCDC50 results in ablation of protein expression across the population, creating a versatile model for investigating gene function in a diverse genetic context.
NCI-H1299 is an established epithelial cell line isolated from a lymph node metastasis of a lung adenocarcinoma. It lacks functional p53 protein due to a homozygous TP53 deletion, making it a widely adopted model for metastatic NSCLC. This p53-null background is particularly relevant for autophagy and NF-??B research, as p53 is a key regulator of both pathways. The cells grow as an adherent monolayer and are highly transfectable, facilitating downstream genetic and pharmacological manipulations.
CCDC50 operates as a dual inhibitor of autophagy and NF-??B signaling. It binds directly to ATG7, obstructing ATG7-mediated LC3 lipidation and autophagosome formation. Simultaneously, CCDC50 associates with IKK??/NEMO, preventing activation of the I??B kinase complex, stabilization of I??B??, and nuclear translocation of p65/RelA. Consequently, CCDC50 suppresses transcription of NF-??B target genes such as Bcl-2 and XIAP. Upstream regulatory inputs include TNF-?? and Toll-like receptor ligands (e.g., LPS), which may signal via NF-??B to modulate CCDC50 expression, forming complex feedback loops.
In the NCI-H1299 background, CCDC50 knockout is expected to derepress autophagic flux and enhance NF-??B-driven transcription, potentially impacting cell survival, apoptosis, and inflammatory cytokine output. This model enables dissection of how CCDC50 loss shifts the balance between pro-survival and pro-death signals in a metastatic NSCLC context. The p53-null status further provides a clinically relevant system to examine p53-independent autophagy and apoptosis regulation, shedding light on mechanisms underpinning tumor progression and therapeutic resistance.
Key applications include monitoring autophagic flux via LC3-II/p62 Western blotting or LC3 immunostaining, assessing NF-??B activity with luciferase reporters or RT-qPCR of Bcl-2/XIAP, and measuring apoptosis/viability by Annexin V/PI flow cytometry or MTS assays. Migration and invasion can be studied using Transwell chambers, while co-immunoprecipitation validates disrupted ATG7/NEMO interactions. Drug sensitivity profiling can reveal synthetic lethal dependencies. For further information, contact Ascent Research.
