The KBTBD4 Knockout SK-HEP-1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population of SK-HEP-1 human hepatic adenocarcinoma cells with targeted disruption of the KBTBD4 gene. This heterogeneous knockout pool contains diverse loss-of-function alleles, providing a robust model to study KBTBD4 function without clonal artifacts. KBTBD4 serves as a substrate adaptor for the Cullin3-RING E3 ubiquitin ligase complex, making this product valuable for investigating ubiquitin-dependent proteasomal degradation in cancer.
The SK-HEP-1 cell line was isolated from ascites of a liver adenocarcinoma patient and is widely employed as a hepatocellular carcinoma model. Molecular profiling indicates an endothelial origin, conferring a hybrid phenotype that facilitates research on tumor cell plasticity and the tumor microenvironment. This background provides a physiologically relevant setting for examining the consequences of KBTBD4 loss.
KBTBD4 bridges the CUL3 scaffold and the RING finger protein RBX1 to recruit substrates for ubiquitination by the CRL3 E3 ligase. Following substrate binding, RBX1 catalyzes ubiquitin transfer from E2 enzymes, tagging proteins for proteasomal degradation. The adaptor function of KBTBD4 is essential for substrate specificity within this complex. Thus, its disruption likely impairs substrate ubiquitination, causing accumulation of undegraded proteins and altered signaling networks.
In SK-HEP-1 cells, KBTBD4 knockout enables investigation of CUL3-RBX1-mediated proteolysis in hepatocellular carcinoma. Though KBTBD4 is implicated in medulloblastoma, its role in liver cancer is poorly defined. This model helps identify KBTBD4-dependent substrates and assess their impact on cell cycle control, apoptosis, and drug sensitivity, revealing potential therapeutic vulnerabilities.
Applications include ubiquitination assays to monitor substrate modification, co-immunoprecipitation for protein interaction analysis, and Western blotting or RT-qPCR to quantify substrate levels. Drug sensitivity profiling with proteasome inhibitors, such as bortezomib, assesses synthetic lethal interactions, while cell cycle and apoptosis assays provide functional readouts. This polyclonal pool also supports cancer dependency mapping screens. For further technical details, please contact Ascent Research.
