The DNAH5 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population that disrupts DNAH5 in human lung adenocarcinoma epithelium. This heterogeneous pool of NCI-H1975 cells harbors diverse Cas9-induced locus disruptions, generating a loss-of-function model for ciliary research without clonal selection. The polyclonal format captures multiple editing outcomes, enabling robust, population-level phenotypic analyses suited for studies of ciliary motility and mucociliary clearance.
NCI-H1975 is a well-characterized NSCLC line from a non-smoker female??s lung adenocarcinoma pleural effusion, carrying activating EGFR exon 19 deletion and mutant TP53. Serum starvation induces primary cilia in these cells, providing a transformed epithelial background for studying ciliary structure and function. This context uniquely permits investigation of ciliary defects alongside oncogenic signaling perturbations.
DNAH5 encodes axonemal dynein heavy chain 5, an outer dynein arm core component that generates ciliary beat force essential for mucociliary clearance. FOXJ1 and RFX3 are key transcriptional regulators, while Notch and Wnt pathways modulate its expression. DNAH5 assembles with DNAI1, DNAI2, DNAL1, and the outer dynein arm docking complex into a functional motor. Knockout of DNAH5 disrupts ciliary motility, abrogating mucociliary transport and potentially deregulating Hedgehog signaling, which depends on intact cilia for Gli transcription factor processing.
DNAH5 knockout in NCI-H1975 recapitulates key primary ciliary dyskinesia features like defective mucociliary clearance and may reveal ciliary roles in cancer cell migration and invasion. These cells?? primary cilia allow direct measurement of ciliary beat frequency, structural integrity, and signaling, linking ciliary dysfunction to Kartagener syndrome and tumor biology. This model bridges genetic ciliopathies and cancer research, enabling dissection of ciliary functions in epithelial homeostasis.
Applications include mechanistic studies of ciliary motility, high-content screening for ciliopathy-modifying compounds, and NSCLC cilia research. Key assays are immunofluorescence for acetylated ??-tubulin and Arl13b, western blotting and RT-qPCR for DNAH5 expression, ciliary beat frequency videomicroscopy, mucociliary clearance assays with fluorescent microspheres, and wound-healing/Transwell migration assays. Hedgehog pathway activity can be assessed with Gli-responsive luciferase reporters. For more information, please contact Ascent Research.
