DNAL1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited human lung adenocarcinoma epithelial cell population with targeted disruption of the DNAL1 gene. This product is supplied as a polyclonal knockout pool, generated by bulk transfection and selection, without clonal isolation. The heterogeneous mixture retains diverse loss-of-function alleles, providing a robust model that mitigates clone-specific artifacts. The polyclonal format is ideal for functional assays requiring population-level representation of gene disruption.
The parental NCI-H1975 cell line harbors EGFR L858R and T790M mutations, representing a well-characterized model of non-small cell lung cancer with acquired resistance to first-generation tyrosine kinase inhibitors. These adherent epithelial cells maintain oncogenic signaling dependencies and are widely employed to study EGFR-targeted therapy, drug resistance, and tumor progression. The combination of an oncogenic driver and a ciliary gene knockout creates a dual perturbation system to explore intersections between cancer biology and ciliary function.
DNAL1 encodes a light chain subunit of the outer dynein arm (ODA) in motile cilia, essential for ciliary beat generation and intraflagellar transport. It functions downstream of master ciliary transcription factors FOXJ1 and RFX, with possible modulation by Notch signaling. DNAL1 assembles into ODA complexes with heavy chains DNAH5 and DNAH9, intermediate chains DNAI1 and DNAI2, and docking components CCDC103 and CCDC114 to anchor the motor to axonemal microtubules. Disruption of DNAL1 abolishes ODA assembly, impairing mucociliary clearance, flow-induced signaling, and ciliary-dependent Hedgehog pathway transduction, which require intact motility and compartmentalized signaling effectors.
In NCI-H1975 cells, DNAL1 knockout mimics defects observed in primary ciliary dyskinesia type 16. Although NCI-H1975 cells are not constitutively ciliated, ciliogenesis can be induced by air-liquid interface culture or serum starvation, enabling functional studies of ciliary motility and signaling. This model permits investigation of how ciliary dysfunction interacts with oncogenic EGFR signals, potentially affecting migration, invasion, and drug sensitivity. The EGFR-mutant background provides a unique platform to examine non-canonical contributions of ciliary proteins to lung adenocarcinoma progression.
Applications include ciliopathy disease modeling, high-content screening for modulators of ciliary motility, and mechanistic studies of dynein arm assembly. Recommended assays encompass Western blotting and RT-qPCR for DNAL1 expression, immunofluorescence for ciliary markers (acetylated tubulin, DNAH5), Sanger sequencing to verify target disruption, video microscopy for ciliary beat frequency, air-liquid interface culture for mucociliary clearance, flow cytometry, and drug sensitivity profiling. For additional information, please contact Ascent Research.
