The KIFAP3 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for studying KIFAP3 gene function. This product features targeted disruption of the KIFAP3 locus in the NCI-H1975 human lung adenocarcinoma epithelial cell line, achieved through CRISPR/Cas9-mediated gene editing, resulting in a heterogeneous pool of cells carrying loss-of-function alleles. The polyclonal format provides a representative population for bulk functional assays without clonal isolation bias, making it suitable for screening and pathway analysis.
NCI-H1975 is a well-characterized epithelial cell line derived from a non-small cell lung adenocarcinoma of a nonsmoking female. These cells endogenously harbor EGFR L858R and T790M mutations, conferring sensitivity and acquired resistance to first-generation tyrosine kinase inhibitors, while retaining wild-type p53 status. As a clinically relevant model for EGFR-mutant NSCLC, NCI-H1975 is widely used to study tumor progression, drug resistance mechanisms, and the tumor microenvironment.
KIFAP3 encodes kinesin-associated protein 3, a non-motor accessory subunit of the heterotrimeric kinesin-2 motor complex. It stabilizes the KIF3A/KIF3B motor complex and mediates cargo binding for anterograde intraflagellar transport (IFT) along microtubules, a process essential for ciliogenesis and cilium-dependent signaling. Transcription is regulated by RFX transcription factors and Sonic hedgehog signaling, while the protein physically interacts with IFT-B complex members such as IFT88 and IFT57, as well as Hook adaptor proteins. KIFAP3-dependent IFT is required for trafficking of ciliary membrane receptors, including Smoothened and Polycystin-1, thereby modulating Hedgehog pathway activity and ciliary membrane composition.
In the NCI-H1975 background, KIFAP3 disruption offers a unique tool to dissect ciliary signaling in EGFR-mutant lung adenocarcinoma. Crosstalk between Hedgehog signaling and oncogenic EGFR pathways may influence tumor growth and resistance, and this knockout model enables studies of how loss of anterograde IFT affects ciliary assembly, Smoothened activation, and downstream transcriptional responses. It provides a platform to probe potential vulnerabilities in drug-resistant cells harboring EGFR L858R/T790M.
This polyclonal knockout model supports a range of applications, including investigation of KIFAP3-dependent ciliogenesis, IFT, and Hedgehog pathway dynamics in cancer biology. Representative assays include Western blotting and RT-qPCR for expression analysis, immunofluorescence staining of ciliary markers (acetylated tubulin, ARL13B), cell proliferation assays, and drug sensitivity testing with erlotinib and osimertinib. Flow cytometry-based phenotyping and Hedgehog reporter assays further facilitate mechanistic studies. For more information, researchers are encouraged to contact Ascent Research.
