The CBR1 Knockout Huh-7 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 hepatocellular carcinoma line, engineered for loss-of-function studies of the CBR1 gene. This heterogeneous polyclonal pool harbors targeted gene disruption across the cell population, enabling robust investigation of CBR1-dependent processes without the clonal selection biases that can arise in monoclonal lines. The knockout model serves as a powerful tool for dissecting carbonyl reductase function in a liver cancer context.
The host Huh-7 cell line was established from well-differentiated hepatocellular carcinoma tissue of a 57-year-old male and is characterized by mutant p53 and wild-type ??-catenin expression. Huh-7 cells are widely used in drug metabolism and toxicity screening, hepatitis virus research, and hepatocarcinogenesis modeling. Their epithelial morphology and retention of key hepatic functions make them a relevant platform for studying redox biology and xenobiotic metabolism in a disease-relevant setting.
CBR1 encodes an NADPH-dependent carbonyl reductase that catalyzes the reduction of endogenous and exogenous carbonyl compounds, including prostaglandins, quinones, and lipid peroxidation products. The enzyme is transcriptionally regulated by NRF2 and AHR in response to oxidative stress and electrophilic compounds. Downstream, CBR1 modulates the levels of prostaglandin E2, menadione, daunorubicin, and 4-hydroxynonenal, directly influencing cellular detoxification and signaling. It functionally interacts with the NADPH cofactor, and its activity can be modulated by flavonoid inhibitors and steroid hormone substrates. CBR1 operates within a network that includes AKR1C enzymes, NQO1, CYP450 oxidoreductases, glutathione S-transferases, COX-2, and prostaglandin E synthase.
In the Huh-7 background, CRISPR-mediated disruption of CBR1 eliminates NADPH-dependent carbonyl reductase activity, leading to impaired detoxification of reactive carbonyl species and altered prostaglandin E2 synthesis. This results in elevated oxidative stress and modified sensitivity to quinone-based chemotherapeutics. The model recapitulates key aspects of hepatocellular carcinoma biology where CBR1 has been implicated in drug resistance and metabolic reprogramming. Consequently, it enables dissection of how carbonyl reductase loss affects tumor cell survival, redox homeostasis, and inflammatory signaling in liver cancer.
The CBR1 Knockout Huh-7 Polyclonal Cells are suited for a range of experimental applications, including investigation of drug metabolism and resistance mechanisms, oxidative stress response pathways, and prostaglandin signaling in hepatocarcinogenesis. Typical readouts involve Western blotting and RT-qPCR for protein and transcript analysis, carbonyl reductase enzymatic activity assays, and MTT/ATP-based viability assays to assess chemosensitivity. Flow cytometry can quantify intracellular ROS levels, while prostaglandin E2 ELISA captures alterations in lipid mediator production. Transcriptomic profiling by RNA-seq and metabolic flux analysis using Seahorse technology provide systems-level insights into the consequences of CBR1 loss. For additional information or to request a quote, please contact Ascent Research.
