The IFT74 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population in which the IFT74 gene has been disrupted to create a loss-of-function model. The polyclonal format provides a heterogeneous pool of edited cells, reflecting a range of loss-of-function alleles that ablate IFT74 protein expression. This product offers a robust platform for studying IFT74-dependent processes without the variability often associated with single-cell clones.
The NCI-H1975 cell line, derived from a pleural effusion of a lung adenocarcinoma patient, is an established model of non-small cell lung carcinoma (NSCLC) carrying the EGFR L858R/T790M double mutation. This genetic background drives oncogenic signaling and confers resistance to first-generation EGFR tyrosine kinase inhibitors. NCI-H1975 cells retain epithelial characteristics and are widely employed to study EGFR-mutant NSCLC biology and therapeutic responses. Importantly, these cells can assemble primary cilia, though the role of ciliary signaling in lung adenocarcinoma remains underexplored.
IFT74 encodes a core subunit of the intraflagellar transport complex B (IFT-B), essential for anterograde trafficking of ciliary cargo along microtubules. IFT74 directly interacts with IFT81, IFT20, IFT27, IFT22, IFT25, IFT54, and cytoplasmic dynein. Its expression is transcriptionally regulated by RFX2, RFX3, FOXJ1, and GLI factors. Disruption of IFT74 impairs primary cilium assembly and cripples cilia-dependent Hedgehog and Wnt signaling. In the Hedgehog cascade, IFT-B components facilitate Smoothened (SMO) translocation into the cilium and subsequent GLI transcription factor activation; in Wnt signaling, ciliary IFT proteins modulate ??-catenin stability and TCF/LEF-mediated transcription.
Within NCI-H1975 cells, IFT74 knockout allows probing of ciliary contributions to oncogenic processes. EGFR signaling cross-talks with Hedgehog and Wnt pathways, and primary cilia may influence these interactions in NSCLC. This knockout model enables investigation of cilia-dependent effects on cell proliferation, migration, drug sensitivity, and epithelial-mesenchymal transition. Additionally, it provides a tool to examine ciliary versus non-ciliary functions of IFT-B components in a clinically relevant lung cancer background.
Typical applications include ciliogenesis assays using immunofluorescence for ciliary markers (acetylated ??-tubulin, ARL13B), Hedgehog reporter assays (GLI-luciferase), and Wnt pathway analysis (TOP/FOP Flash). The cells are suited for proliferation, migration, and invasion studies, as well as transcriptomic profiling by RNA-seq, protein interaction analyses by co-immunoprecipitation, and high-content screening for ciliogenesis modulators. For further information, please contact Ascent Research.
