The IFT140 Knockout NCI-H1975 Polyclonal Cells product comprises a population of NCI-H1975 lung adenocarcinoma cells engineered by CRISPR/Cas9-mediated gene disruption to establish a polyclonal loss-of-function model for intraflagellar transport 140 (IFT140). This polyclonal knockout cell population contains heterogeneous editing events across the target locus, providing a gross functional ablation of IFT140 without clonal selection. The model enables systematic interrogation of ciliary transport and signaling in a defined oncogenic background, supporting mechanistic and drug-discovery studies in non-small cell lung cancer (NSCLC).
The host NCI-H1975 cell line is derived from the pleural effusion of a patient with non-small cell lung adenocarcinoma and harbors activating (L858R) and resistance (T790M) mutations in the epidermal growth factor receptor (EGFR). This dual-mutant background renders cells dependent on EGFR signaling and widely used for investigating acquired resistance to EGFR tyrosine kinase inhibitors. The epithelial morphology and maintained ciliation potential make NCI-H1975 particularly suitable for dissecting cilia-dependent pathways in the context of oncogenic kinase signaling.
IFT140 encodes a core subunit of intraflagellar transport complex A (IFT-A), which mediates retrograde trafficking of cargo within the primary cilium. IFT140 directly interacts with IFT122, IFT121, IFT139, IFT43, and IFT144, coupling to dynein motor proteins for minus-end-directed movement along ciliary microtubules. Disruption of IFT140 impairs ciliogenesis and ciliary maintenance, thereby attenuating Hedgehog signaling by altering the proteolytic processing of GLI transcription factors (GLI1, GLI2, GLI3) downstream of SMO and PTCH1. IFT140 also interfaces with Wnt/??-catenin signaling through its role in ciliary sequestration of pathway components, underscoring its broad regulatory influence.
In the NCI-H1975 EGFR-mutant background, IFT140 knockout allows examination of crosstalk between cilia-dependent pathways and oncogenic EGFR signaling. Primary cilia are increasingly recognized as modulators of drug sensitivity and metastatic behavior in NSCLC, and loss of IFT140-mediated retrograde transport may perturb Hedgehog outputs that influence EGFR inhibitor responsiveness. This polyclonal model thus provides a unique tool to investigate how ciliary dysfunction contributes to the aggressive phenotype of lung adenocarcinoma and to explore therapeutic vulnerabilities arising from combined EGFR and ciliary signaling blockade.
Key applications include dissecting ciliogenesis and Hedgehog signaling in NSCLC, modeling primary cilia dysfunction in acquired drug resistance, and screening for modulators of ciliated cancer cell behavior. Representative experimental approaches encompass western blotting for IFT140 and GLI proteins, quantitative RT-PCR for GLI target genes, immunofluorescence microscopy with acetylated tubulin to assess cilia frequency and length, Shh pathway reporter assays, cell migration analyses, and drug sensitivity testing with EGFR inhibitors. For additional information, please contact Ascent Research.
