The IFT27 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the IFT27 gene, providing a loss-of-function model for studying intraflagellar transport (IFT) and ciliary signaling. As a polyclonal knockout pool, this product contains a diverse set of edited alleles, facilitating functional assays without the clonal selection bias of monoclonal lines.
The parental NCI-H1975 cell line is a widely used human non-small cell lung adenocarcinoma model, established from a female patient. These epithelial cells carry activating EGFR mutations (L858R and T790M), making them instrumental for investigating EGFR-mutant lung cancer and resistance to tyrosine kinase inhibitors. NCI-H1975 cells maintain epithelial morphology and growth factor dependence, serving as a standard platform for oncogenic signaling studies.
IFT27 encodes a small GTPase that functions as an integral subunit of the IFT complex B, a multiprotein assembly essential for anterograde ciliary transport. IFT27 interacts directly with the IFT25/IFT81/IFT74 sub-complex and is regulated by upstream components including the IFT-A complex, BBSome, ARL13B, and guanine nucleotide exchange factors. Through its GTPase cycle, IFT27 controls trafficking of ciliary cargo, including components of the hedgehog signaling pathway. Loss of IFT27 disrupts IFT complex B function, impairing ciliary assembly and the retrograde transport of proteins such as GLI2 and GLI3 transcription factors. This results in defective hedgehog signaling, evidenced by reduced expression of target genes (GLI1, PTCH1) and attenuated ligand responsiveness. IFT27 also influences tubulin polymerization and pericentriolar material organization, linking ciliary dynamics to cell cycle progression.
In NCI-H1975 cells, IFT27 knockout creates a unique tool to dissect the interplay between ciliary signaling and EGFR-driven oncogenesis. Disruption of hedgehog pathway components downstream of IFT27 may alter proliferation, migration, and drug sensitivity of these lung adenocarcinoma cells. Given the emerging role of primary cilia in mediating cellular responses to growth factors and chemotherapeutics, this model enables investigation of cilia-dependent mechanisms that contribute to EGFR TKI resistance. Moreover, it provides a platform to study how ciliary dysfunction associated with ciliopathies such as Bardet-Biedl syndrome or retinal dystrophy may intersect with cancer signaling. By combining a clinically relevant lung cancer background with specific IFT27 deficiency, researchers can interrogate the cell-autonomous functions of intraflagellar transport in tumor maintenance and progression.
Researchers can employ this polyclonal knockout population in a variety of assays to characterize IFT27-dependent phenotypes. Standard applications include western blotting for IFT27 and hedgehog pathway proteins (GLI1, GLI2, SUFU), RT-qPCR of target genes (PTCH1, GLI1), and immunofluorescence for ciliary markers (acetylated tubulin, ARL13B). Functional studies may assess cell proliferation, colony formation, and drug sensitivity to EGFR inhibitors (erlotinib, osimertinib) with phospho-EGFR analysis. Migration and invasion assays can elucidate the impact on metastatic behavior. Multi-omics approaches (RNA-seq, proteomics) can map IFT27-regulated networks, while flow cytometry exploits the heterogeneous pool. For further information, please contact Ascent Research.
