The CCDC14 Knockout NCI-H1975 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 lung adenocarcinoma cell line. This pooled knockout model features targeted disruption of the CCDC14 gene, which encodes a centrosomal protein involved in DNA damage response and centrosome cohesion. The polyclonal format provides a heterogeneous population harboring diverse edits, ensuring a robust loss-of-function system free from clonal artifacts. It enables reliable investigation of CCDC14-dependent processes in a physiologically relevant background.
The NCI-H1975 line is an adherent epithelial cell model established from the metastatic pleural effusion of a female patient with non-small cell lung cancer. These cells carry EGFR L858R and T790M mutations, driving constitutive kinase activity and conferring sensitivity to EGFR-targeted therapies. As a widely used model of EGFR-mutant lung adenocarcinoma, NCI-H1975 cells recapitulate oncogenic signaling, dysregulated proliferation, and DNA repair alterations, providing an apt host for studying CCDC14 function.
CCDC14 is a centrosomal scaffold protein that modulates DNA damage-induced centrosome amplification by binding CEP63 and CEP152, key initiators of centriole duplication. It operates downstream of mitotic and DNA damage kinases, including PLK1, Aurora A, ATM, and ATR, and regulates downstream effectors such as CHK1, CHK2, and p53. CCDC14 also interacts with CDK5RAP2 and PCM1 to coordinate microtubule organization and centrosome cohesion, thereby integrating signals from the ATM-CHK2-p53 and PLK1-CCDC14-centrosome cohesion pathways.
Within the NCI-H1975 context, CCDC14 knockout disrupts centrosome integrity and genome stability, a state compounded by the cell line??s oncogenic EGFR signaling and intrinsic DNA repair dependencies. Loss of CCDC14 is predicted to exacerbate centrosomal abnormalities, leading to mitotic defects and heightened sensitivity to DNA-damaging chemotherapeutics. This model thus serves as a valuable platform to dissect the interplay between centrosomal dysfunction and EGFR-driven tumorigenesis, shedding light on mechanisms of drug resistance in non-small cell lung cancer.
Applications include investigating centrosome biology via immunofluorescence for ??-tubulin, assessing DNA damage responses through western blot of ??H2AX, and performing cell cycle analysis by flow cytometry. The polyclonal knockout cells are also suitable for MTT proliferation assays, drug sensitivity profiling with cisplatin or paclitaxel, and comet assays for DNA damage quantification. These tools support mechanistic studies and translational research. For additional details or custom knockout services, please contact Ascent Research.
