The DLGAP5 Knockout Huh-7 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for studying the microtubule-associated protein DLGAP5 in a human hepatocellular carcinoma background. This product features a polyclonal pool of Huh-7 cells with targeted disruption of the DLGAP5 gene, generating a heterogeneous loss-of-function model. Unlike monoclonal lines, this polyclonal format preserves population-level diversity, which is advantageous for capturing varied phenotypic responses. The knockout model serves as a powerful tool to dissect DLGAP5’s contributions to mitotic spindle assembly, chromosome segregation, and cell proliferation without the need for single-cell clonal isolation.
The host Huh-7 cell line originates from a well-differentiated hepatocellular carcinoma of a human male and is a gold-standard model in hepatocyte biology, liver metabolism, viral hepatitis research, and drug metabolism studies. These epithelial cells retain many liver-specific functions, including expression of hepatic enzymes and transporters, making them highly relevant for investigating liver cancer mechanisms. Their robust proliferative capacity and well-characterized genetic background enable reliable in vitro assays, and they are frequently employed in the study of hepatocarcinogenesis and therapeutic responses.
DLGAP5 functions as a critical regulator of mitotic spindle organization by cooperating with Aurora A kinase and TPX2 to promote bipolar spindle formation and faithful chromosome segregation. It is transcriptionally controlled by E2F1 and FOXM1 and is activated downstream of the Ran GTPase pathway. DLGAP5 directly interacts with Aurora A, TPX2, importin-beta, microtubules, and PLK1, linking it to key mitotic signaling networks. Disruption of DLGAP5 disrupts these interactions, leading to defective spindle assembly, mitotic arrest, and potential induction of apoptosis. As a substrate and partner of Aurora A, DLGAP5 is integral to the Aurora A?CTPX2?Cmicrotubule axis, and its loss perturbs PLK1-mediated checkpoint signaling.
In the context of hepatocellular carcinoma, DLGAP5 is frequently overexpressed and correlates with aggressive disease and poor prognosis. Knockout of DLGAP5 in Huh-7 cells thus offers a clinically relevant model to explore its oncogenic functions and to assess the consequences of its loss on tumor cell viability. The mitotic defects induced by DLGAP5 depletion can be evaluated in a liver cancer setting, providing insights into therapeutic vulnerabilities associated with mitotic spindle dysfunction. This polyclonal knockout population mimics the genetic heterogeneity of tumors, enabling robust assessment of DLGAP5 dependency in hepatic cancer cell proliferation and mitotic fidelity.
Researchers can employ this model for a wide range of applications, including flow cytometric cell cycle analysis, immunofluorescence visualization of aberrant spindle morphology, and functional assays such as MTT-based proliferation, colony formation, and Annexin V apoptosis detection. The cells are suitable for investigating interactions with Aurora A, TPX2, and PLK1, and for conducting loss-of-function screens to identify synthetic lethal partners in liver cancer. By providing a reproducible DLGAP5-deficient background, these polyclonal knockout cells facilitate mechanistic studies and drug discovery efforts targeting the mitotic machinery. For additional technical details and ordering information, please contact Ascent Research.
