The ATOX1 Knockout SK-HEP-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population featuring disruption of the ATOX1 gene. This product is provided as a heterogeneous pool of edited cells, offering a loss-of-function model for studying copper homeostasis and related pathways. By utilizing CRISPR/Cas9-mediated gene disruption in the SK-HEP-1 hepatic adenocarcinoma background, researchers can explore the functional consequences of ATOX1 ablation without the need for single-cell cloning or monoclonal isolation. The polyclonal format maintains a diverse genetic background, which may better represent the complexity of tumor cell populations.
The host cell line, SK-HEP-1, is a human liver adenocarcinoma cell line originally derived from ascitic fluid. It displays an epithelial morphology and is widely employed as a model system for hepatic cancer research. SK-HEP-1 cells have been extensively characterized for their utility in studies of copper metabolism, angiogenesis, and hepatocellular carcinoma biology. Their derivation from a metastatic site renders them particularly suitable for investigating cancer cell migration and invasion mechanisms, traits that are often influenced by copper-dependent enzymes such as lysyl oxidase.
ATOX1 (Antioxidant 1 Copper Chaperone) functions as a cytosolic copper chaperone, binding copper ions and delivering them to the copper-transporting P-type ATPases ATP7A and ATP7B in the trans-Golgi network. This interaction is critical for loading copper into secretory cuproenzymes such as ceruloplasmin and lysyl oxidase. ATOX1 expression is regulated by cellular copper levels and oxidative stress, with the transcription factor NRF2 acting as an upstream mediator. Disruption of ATOX1 via gene knockout impairs copper delivery to ATP7A and ATP7B, leading to reduced cuproenzyme activity, accumulation of cytosolic copper, and elevated oxidative stress.
In the SK-HEP-1 liver adenocarcinoma cell context, ATOX1 knockout allows dissection of copper homeostasis and cancer biology links. Copper supports tumor angiogenesis and proliferation; ATOX1-mediated delivery to lysyl oxidase promotes extracellular matrix remodeling and metastatic spread. This model enables study of disrupted copper trafficking effects on growth, migration, and copper chelator sensitivity. It also serves research into Wilson disease, Menkes disease, and therapeutic strategies targeting copper in cancer.
Typical experimental applications include copper uptake assays using radioactive or stable copper isotopes, quantification of intracellular copper content via ICP-MS, and immunoblotting to assess ATP7A and ATP7B expression levels. Functional assays such as lysyl oxidase activity measurements, ROS detection with DCFDA, MTT proliferation assays, and migration assays are directly supported. This polyclonal knockout population can be used to screen copper chelators for anticancer efficacy or to uncover novel interactions in the copper homeostasis network. For technical details and ordering information, please contact Ascent Research.
