KATNBL1 Knockout NCI-H1975 Polyclonal Cells consist of a population of human NCI-H1975 lung adenocarcinoma cells that have been subjected to CRISPR/Cas9-mediated gene disruption targeting the KATNBL1 locus. This polyclonal knockout cell pool provides a loss-of-function model for investigating the roles of the katanin regulatory subunit B1 (KATNBL1) protein. The CRISPR/Cas9 editing approach generates a heterogeneous mixture of cells with altered KATNBL1 alleles, enabling robust functional studies without clonal selection biases.
The parental NCI-H1975 cell line is derived from a non-small cell lung cancer (NSCLC) patient and harbors activating EGFR mutations (L858R and T790M), which confer sensitivity and acquired resistance to first-generation EGFR tyrosine kinase inhibitors. These adherent epithelial cells serve as a well-established model for studying EGFR-driven lung adenocarcinoma, particularly the T790M-mediated resistance mechanism. The cells display classic epithelial morphology and are widely used in cancer biology, drug discovery, and signal transduction research.
KATNBL1 encodes a non-catalytic regulatory subunit of the katanin microtubule-severing complex. It is phosphorylated by mitotic kinases Aurora A, Plk1, and CDK1, and interacts with the catalytic subunit KATNA1 and ??-tubulin to modulate microtubule dynamics. KATNBL1 is essential for microtubule severing during mitotic spindle assembly, centrosome separation, and chromosome segregation. It functions within an Aurora A?CPlk1?CKATNBL1 axis that governs spindle integrity and fidelity. Disruption of KATNBL1 impairs microtubule remodeling, leading to compromised spindle organization and spindle assembly checkpoint activation. KATNBL1 also participates in ciliogenesis and DNA damage responses.
In NCI-H1975 cells, KATNBL1 loss likely exacerbates microtubule instability, causing mitotic defects and altered cell cycle progression. Because microtubules are critical for intracellular trafficking and signaling, knockout may also affect EGFR trafficking, downstream pathways, and apoptotic thresholds. This model helps dissect how microtubule severing contributes to proliferation and survival of EGFR-mutant NSCLC cells, including those with T790M-mediated resistance. Thus, these polyclonal knockout cells are valuable for studying synthetic lethality, drug resistance, and the interplay between microtubule dynamics and oncogenic signaling.
Typical experimental applications include Western blotting and immunofluorescence to confirm KATNBL1 protein loss and visualize changes in microtubule and mitotic spindle morphology. Functional assays such as cell proliferation, cell cycle analysis by flow cytometry, apoptosis detection, and migration/invasion assays can be employed to assess phenotypic consequences. Drug sensitivity profiling with EGFR inhibitors (e.g., osimertinib, gefitinib) may uncover KATNBL1-dependent vulnerabilities. Live-cell imaging of mitosis provides real-time assessment of spindle assembly and chromosome segregation errors. For further information or custom requests, please contact Ascent Research.
