The CAT Knockout Huh-7 Polyclonal Cells are a polyclonal population derived from the Huh-7 human hepatocellular carcinoma cell line, engineered by CRISPR/Cas9 to disrupt the catalase (CAT) gene. This loss-of-function model generates a cellular environment with impaired hydrogen peroxide detoxification, allowing the study of oxidative stress responses without the biases introduced by clonal selection. The polyclonal format retains a mixture of edited alleles, better reflecting the genetic heterogeneity encountered in tumor biology.
The parental Huh-7 line originates from a well-differentiated liver tumor of a 57-year-old Japanese male and displays epithelial morphology with hepatocyte-like features. Widely used in liver cancer research, Huh-7 cells express liver-specific markers and respond robustly to oxidative agents, making them an ideal host for examining the intersection of redox imbalance and hepatocellular transformation. Their well-characterized growth and metabolic profiles facilitate interpretation of knockout-induced phenotypes.
Catalase (CAT) is a peroxisomal heme enzyme that rapidly converts hydrogen peroxide (H2O2) to water and oxygen, guarding cells against oxidative injury. Its expression is regulated by FOXO transcription factors, NFE2L2 (Nrf2), and PPAR??, and is responsive to insulin/IGF-1 signaling and oxidative stress. CAT functions alongside superoxide dismutase 1 (SOD1) and glutathione peroxidase 1 (GPX1) within the antioxidant network, requiring PEX5 for peroxisomal import and heme as a cofactor. Knockout of CAT leads to H2O2 accumulation, which activates JNK and p38 MAPK kinases, promotes NF-??B signaling, and triggers apoptosis, while also increasing lipid peroxidation and protein carbonylation.
In Huh-7 hepatocellular carcinoma cells, CAT deletion provides a unique tool for dissecting the role of oxidant signaling in tumor progression. The liver’s high metabolic activity creates substantial oxidative challenge, and cancer cells often rewire antioxidant defenses. This model enables investigation of how catalase loss influences proliferation, drug sensitivity, and redox-dependent pathways such as those mediated by NF-??B and MAPK, offering insight into mechanisms of hepatocarcinogenesis and drug-induced hepatotoxicity.
Researchers can employ these cells for oxidative stress research, liver cancer biology, and hepatotoxicity screening. Standard assays include Amplex Red for H2O2 quantification, DCFH-DA flow cytometry for total ROS measurement, western blotting for phospho-JNK and other markers, MTT cell viability testing under H2O2 challenge, and Annexin V/PI apoptosis assays. The model also facilitates antioxidant drug screening and investigation of redox-regulated pathways. For product inquiries, please contact Ascent Research.
