KATNAL1 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. The product offers targeted disruption of the KATNAL1 gene, enabling loss-of-function studies in a defined genetic background. This polyclonal population is suitable for researchers investigating microtubule regulation and mitotic processes in non-small cell lung cancer (NSCLC). The knockout was generated using CRISPR/Cas9 technology to ablate KATNAL1 function without introducing specific described mutations, providing a versatile model for functional genomics.
NCI-H1975 is an epithelial cell line derived from a female non-smoker with lung adenocarcinoma. It harbors activating EGFR mutations L858R and T790M, which are clinically relevant for targeted therapy and acquired resistance. The cells exhibit adherent growth and are widely employed as a NSCLC model system. This genetic context makes them particularly valuable for assessing molecular pathways that intersect with EGFR signaling and microtubule dynamics.
KATNAL1 encodes an ATP-dependent microtubule-severing enzyme, the catalytic subunit of the katanin complex. It functions alongside its regulatory partner KATNB1 to depolymerize microtubules, a process essential for mitotic spindle assembly, chromosome segregation, and ciliogenesis. KATNAL1 activity is regulated by mitotic kinases including Aurora A kinase (AURKA), Polo-like kinase 1 (PLK1), and Cyclin-dependent kinase 1 (CDK1), which phosphorylate KATNAL1 to modulate its severing activity. Downstream, KATNAL1-mediated severing impacts the microtubule cytoskeleton, mitotic spindle morphology, and ciliary axoneme integrity. It interacts with ??-tubulin and various microtubule-associated proteins (MAPs) to target specific microtubule populations. Disruption of KATNAL1 perturbs microtubule homeostasis, leading to defects in cell division and ciliary structure.
In the NCI-H1975 lung cancer context, KATNAL1 knockout likely compromises mitotic progression and may reduce proliferation due to impaired spindle formation. Given the role of microtubules in intracellular trafficking and signaling, the loss of KATNAL1 could sensitize cells to microtubule-targeting chemotherapeutics such as paclitaxel or vinca alkaloids. Moreover, the EGFR mutant background provides a platform to study potential crosstalk between EGFR signaling and microtubule dynamics, and to evaluate whether KATNAL1 deficiency alters responses to EGFR tyrosine kinase inhibitors. This model is thus instrumental for exploring resistance mechanisms and identifying synthetic lethal interactions.
Researchers can employ these polyclonal cells in a variety of assays, including Western blotting and RT-qPCR to confirm KATNAL1 disruption, immunofluorescence staining for microtubule and cilia visualization, flow cytometry for cell cycle and apoptosis analysis, and proliferation assays (e.g., MTT or colony formation). Functional studies may include transwell migration/invasion assays and drug sensitivity testing with paclitaxel or EGFR inhibitors. Live-cell imaging of mitosis can reveal dynamics of spindle assembly defects. The knockout model aids in validating KATNAL1 as a therapeutic target and elucidating its role in NSCLC pathogenesis. For further details and technical support, please contact Ascent Research.
