The KLHL36 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the endogenous KLHL36 gene has been disrupted via CRISPR/Cas9-mediated gene editing. This product provides a heterogeneous pool of edited cells, representing a versatile loss-of-function model for investigating the roles of KLHL36 in hepatocellular carcinoma biology. The polyclonal format captures the diversity of gene edits within the population, facilitating robust downstream applications without clonal selection bias. This knockout model enables systematic exploration of KLHL36-dependent ubiquitin-proteasome pathways and their implications in liver cancer.
The SK-HEP-1 host cell line is a well-characterized human hepatocellular carcinoma cell line originally derived from the ascites of a patient with liver adenocarcinoma. These adherent epithelial cells retain key features of liver cancer cells and are widely used as an in vitro model for studying hepatocarcinogenesis, tumor cell biology, and therapeutic responses. SK-HEP-1 cells provide a relevant background to dissect oncogenic mechanisms related to the ubiquitin-proteasome system, including substrate recognition and protein turnover regulated by Cullin-RING ligases.
KLHL36 functions as a substrate-specific adaptor of the Cullin3-RING E3 ubiquitin ligase (CRL3) complex, where it forms the CRL3 complex with CUL3 and RBX1 to recruit target substrates for ubiquitination. Through its BTB domain, KLHL36 binds CUL3, while its Kelch repeats likely mediate substrate recognition, leading to the transfer of ubiquitin from an E2 ubiquitin-conjugating enzyme to lysine residues on substrates. This polyubiquitination marks proteins for degradation by the 26S proteasome. Although the full repertoire of KLHL36 substrates remains undefined, it is implicated in regulating proteins involved in cell cycle control and stress responses, positioning KLHL36 as a node in proteostatic regulation within hepatocellular carcinoma cells.
In SK-HEP-1 cells, disruption of KLHL36 abolishes the normal function of the CRL3KLHL36 ligase complex, thereby perturbing the ubiquitin-proteasome degradation of its substrate proteins. This provides a powerful tool to dissect the downstream consequences of impaired proteasomal degradation in liver adenocarcinoma cells. The model may reveal how KLHL36 contributes to the turnover of oncogenic or tumor-suppressive factors, offering insights into the molecular pathology of hepatocellular carcinoma where dysregulation of Cullin-RING ligase pathways is frequently observed. Such a system can help elucidate whether KLHL36 acts as a tumor modulator and identify potential vulnerabilities in liver cancer.
Researchers can employ this polyclonal KLHL36 knockout cell population in a wide array of functional assays to investigate the ubiquitin-proteasome system in liver cancer. Representative experiments include Western blotting to assess substrate accumulation, co-immunoprecipitation to validate interactions with CUL3 and RBX1, in vitro ubiquitination assays to monitor E3 ligase activity, and proteasomal degradation assays using inhibitors like MG132. Cell viability assays, such as those measuring responses to chemotherapeutics, can reveal phenotypic consequences. Moreover, siRNA knockdown rescue experiments can confirm gene-specific effects. The model is also suitable for unbiased screens to identify novel KLHL36 substrates via proteomics. For further technical inquiries, please contact Ascent Research.
