The ABCB8 Knockout Huh-7 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population in which the ABCB8 gene has been disrupted in the Huh-7 human hepatocellular carcinoma cell line. This polyclonal pool provides a genetically diverse loss-of-function model suitable for studying ABCB8 biology without clonal selection biases.
Huh-7 is a well-established adherent epithelial cell line derived from a liver tumor of a 57-year-old Japanese male. It serves as a widely used model for hepatocellular carcinoma, hepatocyte function, and viral hepatitis research, offering a hepatic context for investigating iron metabolism, oxidative stress, and drug responses.
ABCB8 encodes a mitochondrial inner membrane transporter that exports iron from the matrix to the intermembrane space, facilitating heme biosynthesis and protecting against mitochondrial iron overload and oxidative stress. Its activity is modulated by iron status, heme, doxorubicin, and the transcription factor NRF2. Downstream, ABCB8 influences mitochondrial iron levels, heme synthesis, ROS production, cytosolic iron-sulfur cluster assembly, and ferroptosis resistance. It interacts with mitochondrial iron-sulfur cluster assembly proteins, heme biosynthetic enzymes, and the chaperone HSPA9. Key pathway components include mitoferrin (SLC25A37), ABCB7, frataxin, ferrochelatase, and IRP1/IRP2. Disruption of ABCB8 leads to mitochondrial iron accumulation and heightened oxidative vulnerability.
In Huh-7 hepatoma cells, ABCB8 knockout creates a relevant model for exploring mitochondrial iron homeostasis in liver cancer. Hepatocytes are central to iron regulation and susceptible to ferroptosis, an iron-dependent cell death implicated in liver disease and cancer therapy. This model allows investigation of ABCB8-dependent modulation of tumor cell survival, drug sensitivity, and metabolic reprogramming, as well as mechanisms of doxorubicin-induced toxicity.
This polyclonal knockout population supports applications in mitochondrial iron homeostasis, heme biosynthesis, and ferroptosis research using assays such as ICP-MS for iron quantification, Western blotting, ROS detection, heme synthesis assays, and cell viability tests under oxidative stress. Additional ferroptosis-focused assays include lipid peroxidation and GPX4 activity measurements. The model is also suited for drug testing and hepatotoxicity studies. For further details, contact Ascent Research.
