The ASB9 Knockout SK-HEP-1 Polyclonal Cells product comprises a CRISPR/Cas9-edited population of polyclonal knockout cells targeting the ASB9 gene in the SK-HEP-1 human liver adenocarcinoma-derived endothelial-like cell line. This polyclonal knockout cell pool provides a loss-of-function model for investigating ASB9 biology, generated through CRISPR/Cas9-mediated gene disruption. As a mixed population, it captures the heterogeneity of editing events, enabling robust functional studies without single-cell cloning bottlenecks.
SK-HEP-1 cells were originally derived from the ascites of a patient with liver adenocarcinoma and exhibit a dual phenotype: they display endothelial morphology and express canonical endothelial markers such as CD31 and von Willebrand factor, yet retain tumorigenic properties and anchorage-independent growth. This unique hybrid character makes SK-HEP-1 a widely used in vitro model for liver sinusoidal endothelium, facilitating research on angiogenesis, tumor microenvironment interactions, and hepatic drug transport.
ASB9 functions as a substrate-recognition subunit within the Cullin 5?CRbx2?CElongin BC (CRL5) E3 ubiquitin ligase complex. Upon activation by upstream cytokine signals, including IL-6 and IFN-?? via the JAK-STAT cascade, ASB9 recruits specific substrates such as mitochondrial creatine kinase (CKMT1) for ubiquitination. The ubiquitinated targets are subsequently degraded by the 26S proteasome, thereby attenuating JAK-STAT-mediated transcription. ASB9 directly interacts with Elongin B, Elongin C, Cullin 5, and Rbx2, linking cytokine stimulation to proteasomal degradation and serving as a critical regulator of cellular inflammatory responses and protein homeostasis.
In the SK-HEP-1 background, ASB9 knockout disrupts this negative-feedback arm of JAK-STAT signaling, potentially leading to sustained pathway activation and altered downstream endothelial functions. Given the cell line??s dual liver cancer and endothelial identity, this model is uniquely suited to dissect how ASB9-mediated ubiquitination shapes cytokine responses in hepatic tumor microenvironments, influences angiogenic signaling, and contributes to metabolic reprogramming. The polyclonal knockout approach avoids artifacts associated with single-cell selection, preserving population-level biological variability.
Researchers can employ this polyclonal knockout cell pool in a wide array of functional assays. Co-immunoprecipitation and ubiquitination assays facilitate mapping of ASB9?Csubstrate interactions, while phospho-STAT flow cytometry and RNA-seq enable profiling of JAK-STAT pathway dynamics. Proteasome activity and cell proliferation/apoptosis assays assess the impact on protein turnover and cell fate, and metabolomics or migration assays explore consequences for metabolic and endothelial behavior. This product is thus a versatile tool for investigating ubiquitin-proteasome system dysregulation in liver cancer, cytokine signaling, and immune modulation. For further information, please contact Ascent Research.
