The CBR3 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population of Huh-7 hepatocellular carcinoma cells. This heterogeneous pool harbors CBR3 gene disruptions from non-clonal editing, providing a bulk loss-of-function model. As a pooled knockout product, it avoids clonal artifacts and enables robust study of CBR3-dependent carbonyl reduction in hepatic contexts.
The Huh-7 parental line is a well-differentiated hepatocyte-derived carcinoma from a 57-year-old Japanese male, retaining hepatocyte-like properties and drug-metabolizing enzyme expression. It is a standard model for hepatitis C virus, drug metabolism, and liver cancer research, and its oxidative stress responsiveness makes it ideal for CBR3 detoxification studies.
CBR3 encodes an NADPH-dependent carbonyl reductase that metabolizes endogenous and xenobiotic carbonyls, such as prostaglandins and 4-oxo-2-nonenal, protecting against oxidative damage. Its expression is under NFE2L2 (Nrf2) transcriptional control, normally inhibited by KEAP1. Upon oxidative stress, Nrf2 drives CBR3 alongside NQO1, HMOX1, and GCLC. CBR3 cooperates with NQO1 and GSTP1 in reactive carbonyl reduction; its knockout leads to carbonyl accumulation and oxidative stress, eliciting compensatory Nrf2 responses.
In the hepatic setting, CBR3 loss is highly relevant to hepatocellular carcinoma biology and drug handling. The liver??s central role in xenobiotic detoxification involves CBR3 in inactivating drugs like doxorubicin, linking it to anthracycline cardiotoxicity and chemoresistance. Eliminating CBR3 activity in Huh-7 cells permits dissection of carbonyl-reducing pathways that determine drug sensitivity and oxidative liver injury, and allows exploration of how CBR3 deficiency modulates Nrf2-dependent antioxidant programs under chronic oxidative conditions typical of HCC.
This polyclonal knockout pool supports various applications: investigating detoxification pathways via carbonyl reductase assays and western blotting; measuring oxidative stress with DCFDA and TBARS; assessing drug sensitivity by MTT assay after doxorubicin exposure; and screening Nrf2 activators with ARE luciferase reporters and RT-qPCR for NQO1 and HMOX1. It is a valuable tool for drug-induced liver injury, chemoresistance, and redox biology research. For technical assistance, contact Ascent Research.
