The IFT20 Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population targeting the IFT20 gene in the human lung adenocarcinoma epithelial cell line NCI-H1975. This polyclonal population, derived from Homo sapiens, features broad disruption of IFT20 across a heterogeneous pool of edited cells, enabling robust loss-of-function studies without clonal selection bias. The product offers researchers a physiologically relevant system to investigate IFT20-dependent processes in a genetically characterized non-small cell lung cancer (NSCLC) background.
The host NCI-H1975 cell line originates from a non-smoking female patient with lung adenocarcinoma and harbors activating EGFR L858R and secondary T790M mutations, rendering it an established model for EGFR-targeted therapy resistance in NSCLC. These epithelial cells retain key properties of lung adenocarcinoma, including reliance on oncogenic EGFR signaling and responsiveness to kinase inhibitors, making them a clinically pertinent platform for dissecting tumor biology. The integration of IFT20 knockout into this EGFR-mutant context provides a unique tool to explore crosstalk between ciliary signaling and oncogenic pathways.
IFT20 encodes an intraflagellar transport (IFT) protein that is a core component of IFT complex B, essential for anterograde ciliary trafficking and ciliogenesis. It functions in Golgi-to-cilium protein delivery and immune synapse formation, interacting with complex B partners such as IFT52, IFT88, and IFT57, as well as the ciliary membrane protein SMO and the PDGF receptor PDGFR??. IFT20 expression is transcriptionally regulated by RFX transcription factors and FOXJ1, and its activity mediates signaling downstream of Hedgehog and Wnt pathways, affecting downstream effectors like GLI1 and PTCH1. By physically associating with GMAP210 and exocyst complex components, IFT20 coordinates ciliary protein sorting and membrane trafficking.
In NCI-H1975 cells, IFT20 knockout disrupts primary cilium assembly and impairs ciliary-dependent signal transduction, including Hedgehog and Wnt cascades that are increasingly implicated in lung adenocarcinoma progression. Loss of ciliary function may alter cellular responses to EGFR inhibition, modify proliferative and migratory behavior, and shift differentiation states, providing a model to study how ciliary defects intersect with oncogenic drivers. This polyclonal knockout system is particularly valuable for examining context-dependent roles of IFT20 in EGFR-mutant NSCLC, where cilia status may influence drug sensitivity and metastatic potential.
Researchers can apply this knockout model to a range of studies: immunofluorescence using cilia markers acetylated tubulin or ARL13B to assess ciliogenesis; Western blotting to confirm IFT20 ablation; RT-qPCR for Hedgehog target genes GLI1 and PTCH1; RNA-seq transcriptomic profiling; cell proliferation assays; drug sensitivity testing with EGFR inhibitors like osimertinib; and migration/invasion assays to evaluate metastatic behavior. The polyclonal nature supports population-level analyses while avoiding clonal artifacts. For detailed protocols or batch-specific characterization, please contact Ascent Research.
