The DLGAP5 Knockout NCI-H1975 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the DLGAP5 gene has been disrupted, creating a loss-of-function model for mitotic regulation studies. This product provides a heterogeneous population of NCI-H1975 cells carrying diverse allele edits, enabling functional investigation of DLGAP5 without clonal isolation artifacts. The polyclonal format offers a robust and reproducible platform for probing the immediate consequences of DLGAP5 ablation on spindle assembly and chromosome segregation.
The parental NCI-H1975 cell line is a widely employed human lung adenocarcinoma model derived from a non-small cell lung cancer (NSCLC) patient. It harbors endogenous activating mutations in EGFR (L858R and T790M), rendering it a clinically relevant system for studying EGFR-driven oncogenesis and therapeutic resistance. These cells retain epithelial characteristics and are amenable to standard culture, genetic manipulation, and high-content imaging, making them ideal for examining mitosis in a lung cancer context.
DLGAP5 (Discs Large-Associated Protein 5) is a microtubule-associated protein crucial for mitotic spindle integrity. It localizes to kinetochore fibers and stabilizes them to promote chromosome congression and alignment. Mechanistically, DLGAP5 operates downstream of Aurora A kinase, which phosphorylates it, and upstream of TPX2 and the kinesin motor Eg5 (KIF11), facilitating their recruitment to spindle microtubules. Transcriptional control by E2F factors and FoxM1 links DLGAP5 expression to cell cycle entry. Additionally, DLGAP5 interacts with importin beta and contributes to spindle assembly checkpoint signaling through modulation of BubR1 and Mad2, thereby ensuring accurate chromosome segregation. This positions DLGAP5 at the intersection of Aurora A/PLK1 signaling and kinetochore regulation.
In the NCI-H1975 background, DLGAP5 disruption interrogates the dependency of EGFR-mutant lung adenocarcinoma on mitotic fidelity. DLGAP5 overexpression is associated with poor prognosis in NSCLC and hepatocellular carcinoma, suggesting that its function may be exploited by rapidly proliferating cancer cells. The polyclonal knockout population permits assessment of how loss of DLGAP5 impacts spindle morphology, cell cycle progression, and sensitivity to mitotic inhibitors such as Aurora A kinase inhibitor MLN8237. This model thereby bridges oncogenic signaling and mitotic vulnerability in a genetically defined lung cancer context.
This knockout product is suited for diverse experimental applications including western blotting to confirm DLGAP5 loss, immunofluorescence microscopy to visualize aberrant spindle formation, flow cytometry for cell cycle analysis, and live-cell imaging to track mitotic delays. Functional assays such as colony formation, apoptosis measurement, and drug sensitivity profiling with MLN8237 or other spindle poisons can elucidate the role of DLGAP5 in tumor cell fitness. RNA-sequencing studies can further reveal transcriptomic changes upon DLGAP5 depletion. These cells provide a versatile tool for both mechanistic dissections and preclinical drug testing in lung cancer. For additional details or to inquire about custom services, please contact Ascent Research.
