The CCDC22 Knockout NCI-H1299 Polyclonal Cells product consists of a heterogeneous pool of NCI-H1299 cells that have undergone CRISPR/Cas9-mediated disruption of the endogenous CCDC22 locus, yielding a polyclonal loss-of-function model. This population retains the genetic diversity inherent to polyclonal editing and is suitable for bulk functional assays where clonal selection is not required. The knockout disrupts CCDC22 protein expression, enabling researchers to interrogate the role of CCDC22 in endosomal trafficking, signaling, and disease-related processes.
The host cell line, NCI-H1299, is a widely used human non-small cell lung carcinoma (NSCLC) line established from a lymph node metastasis of a lung adenocarcinoma. It is characterized by a homozygous deletion of TP53, rendering it p53-null, and it displays an epithelial morphology. NCI-H1299 serves as a robust model for studying metastatic potential, drug resistance, and signal transduction in lung cancer, making it an informative background for investigating tumor suppressor and oncogenic pathways.
CCDC22 encodes a core subunit of the CCC (COMMD-CCDC22-CCDC93) complex, which cooperates with the WASH and retromer complexes to mediate endosomal recycling. It stabilizes COMMD proteins, particularly COMMD1, and forms a structural backbone with CCDC93. This complex is critical for recycling the copper transporters ATP7A and ATP7B, thus maintaining copper homeostasis, and it negatively regulates NF-??B signaling via COMMD1-dependent processes. Upstream regulators RAB5 and RAB7 direct endosomal localization of CCC. Downstream, CCDC22 knockout leads to COMMD protein destabilization, defective ATP7A/B trafficking, and dysregulated NF-??B activity, as reflected by altered IKK??/IKK?? phosphorylation and p65 nuclear translocation.
In NCI-H1299 p53-null lung adenocarcinoma cells, CCDC22 knockout offers a model to study the crosstalk between copper metabolism and NF-??B signaling in cancer. Both pathways are linked to tumor progression and drug resistance, and the absence of p53 heightens dependence on alternative regulatory mechanisms. This polyclonal knockout population allows functional investigation of the CCC complex in a metastatic NSCLC context without clonal bias.
This polyclonal knockout model is well-suited for functional studies of endosomal recycling, copper homeostasis, and NF-??B signaling. Experimental approaches include copper uptake/efflux assays with radioactive or fluorescent probes, NF-??B luciferase reporter assays, drug sensitivity screens using copper chelators (e.g., tetrathiomolybdate) or IKK inhibitors, and confocal immunofluorescence to track ATP7A/B localization. Western blotting and RT-qPCR can quantify COMMD1 stability and NF-??B target gene expression. For technical support or custom requests, please contact Ascent Research.
