The EFCAB14 Knockout NCI-H1975 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the EFCAB14 gene in the human NCI-H1975 lung adenocarcinoma cell line. As a polyclonal pool, this product contains a diverse collection of edited cells harboring individual gene disruptions generated by CRISPR/Cas9-mediated targeting, facilitating loss-of-function studies without the need for clonal isolation. This model is specifically tailored for investigations into ciliary biology, calcium-mediated signaling, and tumor cell dynamics.
The NCI-H1975 host cell line is a well-characterized human lung adenocarcinoma epithelial line derived from a non-smoking female patient. It harbors the activating EGFR L858R mutation and the secondary EGFR T790M gatekeeper mutation, conferring resistance to first-generation EGFR tyrosine kinase inhibitors (TKIs). This mutational background makes NCI-H1975 an essential model for studying acquired TKI resistance mechanisms and for preclinical evaluation of next-generation EGFR inhibitors in non-small cell lung cancer (NSCLC).
EFCAB14 is a calcium-binding EF-hand protein that functions as a molecular calcium sensor, stabilizing microtubule structures essential for ciliogenesis and flagellar motility. The protein directly interacts with tubulin and ciliary trafficking proteins IFT88 and SPAG6, and its activity is modulated by cytosolic calcium concentrations and phosphorylation by unknown kinases. Within the ciliogenesis cascade, EFCAB14 integrates calcium signals with intraflagellar transport (IFT) machinery, including the IFT-A and IFT-B complexes, the BBSome, and motor proteins kinesin-2 and cytoplasmic dynein 2, to regulate the assembly and maintenance of the ciliary axoneme.
Disruption of EFCAB14 in NCI-H1975 cells is expected to impair primary cilium formation and calcium-dependent cytoskeletal dynamics, which may compromise cilium-dependent Hedgehog signaling and promote a more aggressive, drug-resistant phenotype. Given the emerging role of primary cilia in modulating oncogenic signaling, metabolic adaptation, and therapeutic resistance, this knockout model provides a unique platform to dissect the contribution of ciliary dysfunction to EGFR TKI resistance in lung adenocarcinoma. It can be used to investigate whether ciliary loss sensitizes or desensitizes tumor cells to targeted therapies and to explore cilia-mediated regulation of the tumor microenvironment.
This polyclonal knockout cell pool is compatible with a wide array of experimental approaches: Western blotting to verify EFCAB14 protein depletion; immunofluorescence staining for ciliary markers acetylated tubulin and ARL13B to assess ciliogenesis efficiency; RT-qPCR profiling of ciliary gene expression; live-cell calcium imaging to monitor intracellular calcium dynamics; cell migration and invasion assays; and EGFR TKI dose?Cresponse or apoptosis assays. Additionally, the model can be employed in synthetic lethality screens with EGFR inhibitors, high-content imaging studies, and co-culture systems to evaluate tumor?Cstromal crosstalk. For detailed product inquiries, please contact Ascent Research.
