The CCDC127 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population engineered for targeted disruption of the CCDC127 gene in the NCI-H1975 human lung adenocarcinoma cell line. This polyclonal knockout product provides a heterogeneous pool of edited cells, enabling loss-of-function studies of CCDC127 without clonal selection biases. The cells were generated via CRISPR/Cas9-mediated gene disruption, yielding a mixed allelic population that collectively abolishes functional protein expression.
The NCI-H1975 host cell line was isolated from a pleural effusion of a female non-small cell lung cancer (NSCLC) patient and displays adherent epithelial morphology. It carries both EGFR L858R and T790M mutations, which drive constitutive kinase activity and confer resistance to first-generation EGFR tyrosine kinase inhibitors. This well-characterized NSCLC model is widely used to study drug resistance mechanisms, signaling crosstalk, and tumor progression.
CCDC127 is a centriolar satellite protein that localizes to the centrosome and basal body, interacting with PCM1 and other satellite components to facilitate primary cilium assembly. The primary cilium is essential for Hedgehog signaling; CCDC127 thus enables pathway activation by promoting ciliary trafficking of IFT proteins and proper transduction through SMO, PTCH1, and the GLI transcription factors. Loss of CCDC127 impairs ciliogenesis, leading to attenuated Hedgehog target gene expression and disruption of downstream cellular responses.
In the NCI-H1975 background, CCDC127 knockout may uncover crosstalk between Hedgehog signaling and EGFR-driven oncogenic pathways. As primary cilia can influence drug sensitivity and cell cycle regulation, CCDC127 loss could modulate responses to EGFR inhibitors, thereby impacting therapeutic resistance. The EGFR L858R/T790M mutant context provides a clinically relevant system for investigating how centrosomal proteins contribute to NSCLC maintenance and acquired resistance.
Researchers can employ this knockout model to assess ciliogenesis via immunofluorescence for acetylated tubulin or ARL13B, quantify Hedgehog pathway activation by RT-qPCR of GLI1 or PTCH1, and confirm protein depletion by Western blotting. Functional assays such as drug sensitivity testing with gefitinib or osimertinib, cell cycle analysis, apoptosis assays, and migration/invasion assays further elucidate CCDC127??s role in NSCLC biology. For additional information, please contact Ascent Research.
