The CCDC25 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting the CCDC25 gene in the human lung adenocarcinoma line NCI-H1299. This pool of gene-edited cells enables investigation of CCDC25 function without the clonal selection bias inherent in single-cell-derived knockout lines. The polyclonal format is well-suited for pooled functional studies, providing a representative range of editing outcomes to analyze CCDC25-dependent phenotypes at the population level.
NCI-H1299 is a p53-deficient cell line derived from a lymph node metastasis of lung adenocarcinoma. Its metastatic origin and p53-null status mimic aggressive cancer characteristics, including heightened migratory potential and altered stress signaling. This background is thus ideal for examining genes involved in late-stage tumor progression and metastatic dissemination, offering a relevant model for lung cancer biology.
CCDC25 encodes a receptor for DNA associated with neutrophil extracellular traps (NETs), transducing signals that drive cancer cell migration and metastasis. Upon NETs-DNA binding, CCDC25 activates integrin-linked kinase (ILK) and the adaptor ??-parvin, triggering the small GTPases RAC1 and CDC42 to remodel the actin cytoskeleton. This NETs-CCDC25-ILK-??-parvin-RAC1/CDC42 axis enhances cell motility and invasion. Additionally, CCDC25 serves as a cytoplasmic DNA sensor, triggering NF-??B activation via IKK complex interactions, thus linking extracellular NETs cues and intracellular DNA sensing to pro-metastatic and inflammatory responses.
Knocking out CCDC25 in p53-null, metastasis-derived NCI-H1299 cells is expected to attenuate responses to NETs and disrupt the ILK-??-parvin-RAC1/CDC42 pathway, blunting migratory and invasive capabilities. This model is directly relevant for dissecting NET-mediated signaling in aggressive lung adenocarcinoma. Moreover, the p53 deficiency ensures that DNA-sensing and NF-??B pathways can be studied without interference from p53-dependent apoptosis or senescence, providing a clear platform for analyzing CCDC25-driven signaling.
These polyclonal knockout cells are suitable for Transwell migration and invasion assays, wound healing studies, and in vivo metastasis experiments to evaluate the contribution of CCDC25 to tumor dissemination. Biochemical assays, including co-immunoprecipitation of the CCDC25-ILK complex, Western blotting for downstream targets, and NF-??B reporter assays, allow detailed mechanistic dissection. Immunofluorescence can assess NETs binding, and RAC1/CDC42 activity assays can verify pathway activation. The model supports research on cancer metastasis, NETosis, and innate DNA sensing. For further information, please contact Ascent Research.
