The CCDC127 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of the human NCI-H1299 lung adenocarcinoma cell line, designed to disrupt the CCDC127 gene and eliminate functional expression of the encoded centrosomal protein. This polyclonal pool comprises multiple independent gene-disruption events generated by CRISPR/Cas9-mediated targeting, avoiding the clonal biases inherent in single-cell-derived knockouts while preserving the heterogeneous genetic background that more closely mirrors polyclonal tumor cell populations. The product provides a robust and versatile loss-of-function model for investigating CCDC127-dependent cellular processes in a well-characterized cancer cell background.
NCI-H1299 is an epithelial cell line derived from a lymph node metastasis of a non-small cell lung adenocarcinoma. These cells carry a homozygous deletion of the TP53 tumor suppressor gene, rendering them p53-null and deficient in p53-mediated cell cycle arrest and apoptosis. Consequently, NCI-H1299 exhibits unchecked proliferation, chromosomal instability, and metastatic propensity, making it a widely employed model for studying advanced lung cancer biology, evaluating therapeutic agents, and probing mechanisms that compensate for p53 loss in malignant progression.
CCDC127 encodes a coiled-coil domain-containing protein that localizes to centrosomes, where it functions as a scaffold involved in microtubule organization, centrosome duplication, and cell cycle progression at the G2/M transition. It operates within a molecular network centered on the centriole biogenesis pathway: the kinase PLK4 is recruited by CEP152 and CEP192, which together facilitate the phosphorylation of STIL and the loading of SAS?6, essential steps for procentriole assembly. CCDC127 is proposed to interact with centrosomal scaffold proteins and modulate downstream microtubule nucleation factors??including ???tubulin ring complex components??and pericentriolar material proteins, thereby ensuring proper spindle pole formation and mitotic spindle integrity. Although its direct upstream regulation remains unknown, CCDC127 is likely controlled by cell cycle-dependent kinases that govern centrosome maturation.
In the context of p53-deficient NCI-H1299 cells, CCDC127 loss-of-function provides a powerful system to examine how centrosome dysfunction exacerbates genomic instability and aneuploidy. The absence of p53 checkpoints forces reliance on mitotic fidelity for survival; consequently, disruption of centrosome homeostasis can unmask synthetic lethal interactions or reveal vulnerabilities exploitable by microtubule-targeting agents. This model is particularly suited for dissecting the interplay between centrosome defects and cancer cell behavior, including proliferation, migration, and drug resistance, without confounding p53-dependent stress responses.
Researchers can deploy this KO pool in a variety of experimental paradigms: immunofluorescence staining to visualize centrosome number and microtubule architecture, western blotting to confirm CCDC127 protein depletion, flow cytometry for detailed cell cycle profiling, and proliferation/viability assays to assess growth defects. Mitotic index quantification, microtubule regrowth assays to evaluate nucleation capacity, and RNA?seq transcriptomic analysis further enable comprehensive phenotypic characterization of centrosomal pathway disruption. For further information or technical support, contact Ascent Research.
