The CBR3 Knockout 786-O Polyclonal Cells product comprises a pool of 786-O cells that have been subjected to CRISPR/Cas9-mediated gene disruption targeting the CBR3 locus. This polyclonal knockout cell population is not a clonal line; rather, it represents a heterogeneous mixture of cells carrying diverse loss-of-function edits within the CBR3 gene, resulting in abrogation of carbonyl reductase 3 (CBR3) enzymatic activity. The product serves as an investigative tool for probing CBR3-dependent biological processes without the confounding effects of clonal selection. It is suitable for applications where a polyclonal pool best models population-level responses to genetic perturbation, including studies of drug metabolism, detoxification pathways, and cancer cell behavior.
The host cell line, 786-O, is a well-characterized model of clear cell renal cell carcinoma (ccRCC). Originating from a primary human renal adenocarcinoma, 786-O cells are deficient in the von Hippel?CLindau (VHL) tumor suppressor protein, leading to constitutive stabilization and activation of hypoxia-inducible factors HIF-1?? and HIF-2?? under normoxic conditions. This VHL loss induces a pseudohypoxic phenotype that drives metabolic reprogramming, angiogenesis, and tumor progression. The malignant epithelial nature of 786-O cells makes them particularly relevant for research into ccRCC pathogenesis, drug resistance mechanisms, and hypoxia-related signaling. The introduction of a CBR3 knockout in this genetic background permits dissection of carbonyl reduction pathways in a context that mimics the redox and metabolic abnormalities of renal carcinoma.
CBR3 encodes an NADPH-dependent short-chain dehydrogenase/reductase that catalyzes the reduction of a broad range of carbonyl substrates, including xenobiotic quinones (such as the anthracycline doxorubicin), lipid peroxidation?Cderived reactive aldehydes (e.g., 4-hydroxynonenal), and intermediates in prostaglandin metabolism. The CBR3 enzyme functions within a network regulated by the NFE2L2/Nrf2 pathway; under oxidative stress or electrophilic challenge, NFE2L2 is released from KEAP1-mediated repression and drives transcription of CBR3. Downstream, CBR3 generates reduced metabolites like doxorubicinol and detoxified aldehyde species, thereby modulating cellular responses to chemotherapeutic agents and oxidative damage. CBR3 also participates in arachidonic acid metabolism, influencing prostaglandin F2?? levels. NADPH serves as an essential cofactor, and potential interactions with other carbonyl reductases (e.g., CBR1) may further shape the metabolic fate of substrates. In the knockout pool, disruption of CBR3 is predicted to impair these NADPH-dependent reductase activities, leading to accumulation of reactive carbonyls and altered drug sensitivity.
Within the 786-O ccRCC context, CBR3 knockout addresses a critical gap in understanding how carbonyl detoxification influences tumor cell fitness and drug response. The VHL-deficient, pseudohypoxic environment of 786-O cells is characterized by elevated reactive oxygen species and altered metabolic flux; CBR3 normally mitigates oxidative stress by reducing harmful aldehydes. Its loss may therefore sensitized these cells to anthracycline-induced cytotoxicity or other carbonyl-generating stressors. Additionally, since ccRCC frequently exhibits chemoresistance, this model can be employed to explore whether CBR3 contributes to resistance mechanisms or whether its absence reveals synthetic lethalities. The polyclonal nature of the knockout population allows researchers to observe the overall impact of CBR3 ablation without clonal artifacts, making it ideal for population-based assays of viability, apoptosis, and drug metabolism.
Researchers can apply this polyclonal knockout model to a variety of experimental paradigms. Common applications include drug resistance studies, where the effect of CBR3 loss on doxorubicin or daunorubicin cytotoxicity can be quantified using MTT viability assays or Annexin V/PI apoptosis assays; metabolic tracing of anthracycline conversion via HPLC-based doxorubicin metabolism assays; and investigations into reactive oxygen species (ROS) handling using DCFDA probes. The product is also suited for carbonyl reductase activity assays, biomarker validation, and dissection of NFE2L2/KEAP1 pathway signaling. Expression-level confirmation can be performed by Western blotting or RT-qPCR. These tools enable a comprehensive analysis of CBR3 function in renal carcinoma biology. For additional details, technical specifications, or support, please contact Ascent Research.
