The CBR4 Knockout Huh-7 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human hepatocellular carcinoma Huh-7 cell line, engineered to disrupt the CBR4 gene. This polyclonal pool provides a heterogeneous loss-of-function model for investigating CBR4-dependent processes in a hepatic epithelial background. The gene disruption targets the genomic locus encoding the NADPH-dependent carbonyl reductase CBR4, generating a versatile tool for studying metabolic and detoxification pathways without clonal effects.
Huh-7 is a well-differentiated, epithelial hepatocellular carcinoma cell line isolated from a 57-year-old Japanese male liver tumor. It serves as a widely used model for hepatic metabolism, drug-induced liver injury, and hepatocellular carcinoma biology, retaining key liver functions including phase I and II metabolism. Its adherent morphology and robust growth facilitate diverse in vitro assays, making this knockout model physiologically relevant for studying CBR4 in liver-derived cells.
CBR4 encodes an NADPH-dependent carbonyl reductase that reduces carbonyl substrates to alcohols, modulating the metabolism of endogenous and exogenous compounds. Regulated by NRF2 under oxidative stress and electrophile exposure, CBR4 reduces 15-keto-prostaglandin E2 to 13,14-dihydro-15-keto-PGE2, and detoxifies xenobiotic carbonyls, including CYP450-generated reactive intermediates. The enzyme uses NADPH as a cofactor and interacts with prostaglandin substrates to control signaling and redox balance. Thus, CBR4 integrates oxidative stress responses with xenobiotic and endobiotic metabolism.
Disruption of CBR4 in Huh-7 cells offers a relevant platform to probe carbonyl metabolism in liver cancer. This model allows examination of how loss of prostaglandin and xenobiotic reduction impacts proliferation, chemosensitivity, and metabolic adaptation. Given the liver’s detoxification role, it is particularly suited for investigating the interplay between carbonyl reductase activity, chemoresistance, and oxidative damage, illuminating potential therapeutic targets in hepatocellular carcinoma.
These polyclonal knockout cells support applications such as enzyme activity assays to confirm loss of function, Western blotting and RT-qPCR for target validation, and drug sensitivity testing (e.g., cisplatin, doxorubicin) to assess chemoresistance. Metabolic profiling elucidates changes in arachidonic acid metabolism and xenobiotic clearance, while ROS detection quantifies oxidative stress. The polyclonal nature is advantageous for population-level studies, screening, and functional genomics. For further information, please contact Ascent Research.
