The DLGAP5 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population generated from the SK-HEP-1 human hepatocellular carcinoma line by targeted disruption of the DLGAP5 gene. This loss-of-function model enables investigation of DLGAP5-dependent phenotypes within a hepatic cancer context. The polyclonal format captures a heterogeneous pool of edited genotypes, minimizing clonal selection bias and supporting population-level analyses of proliferation, cell cycle dynamics, and mitotic fidelity.
The SK-HEP-1 cell line is an adherent epithelial model originally derived from ascites fluid of a liver adenocarcinoma patient. It is widely employed as a hepatocellular carcinoma system, retaining key hepatic tumor characteristics for applications in cancer biology, drug screening, and signaling studies. Its metastatic origin renders it particularly suitable for studying aggressive cancer behaviors such as migration and invasion, offering a physiologically relevant platform for evaluating DLGAP5 knockout effects.
DLGAP5 is a critical mitotic spindle assembly factor that stabilizes kinetochore-microtubule attachments to ensure faithful chromosome segregation. It is phosphorylated and activated by Aurora A kinase, functioning downstream of Aurora A signaling, and is transcriptionally regulated by E2F factors and the PI3K/AKT pathway. DLGAP5 directly interacts with TPX2, INCENP, survivin, tubulin, and kinesin-5 to drive microtubule stabilization and proper chromosome alignment. Overexpressed in hepatocellular, gastric, breast, lung, and colorectal cancers, DLGAP5 promotes tumor cell proliferation by enforcing mitotic progression, making it an attractive therapeutic target.
Within the SK-HEP-1 hepatocellular carcinoma model, DLGAP5 knockout provides a unique tool to dissect its mitotic contributions to liver cancer cell survival. Given the frequent dysregulation of Aurora A and PI3K/AKT pathways in this cancer type, this knockout system enables studies of synthetic lethal interactions and the consequences of impaired kinetochore-microtubule dynamics, including chromosome instability and aneuploidy. It thereby facilitates identification of context-specific vulnerabilities exploitable by therapeutic intervention.
Researchers can utilize this polyclonal knockout model in a diverse array of assays, including western blotting to verify DLGAP5 ablation, immunofluorescence to assess mitotic spindle morphology, and flow cytometry for cell cycle and ploidy analysis. Functional studies such as proliferation, colony formation, migration, and invasion assays quantify tumorigenic properties, while apoptosis and phospho-signaling profiling (e.g., Aurora A substrate phosphorylation) reveal downstream signaling alterations. RNA-sequencing offers transcriptome-wide insights, and drug sensitivity screening with mitotic inhibitors (Aurora A, kinesin-5 inhibitors) validates DLGAP5 as a druggable node. For further information, please contact Ascent Research.
