The CAT knockout HEK293T polyclonal cells are a CRISPR/Cas9-edited polyclonal population with targeted disruption of the catalase (CAT) gene. Generated in the widely used HEK293T background, this loss-of-function model enables investigation of catalase-dependent redox mechanisms. Its polyclonal nature ensures a heterogeneous allele mixture, allowing robust gene function assessment without clonal selection artifacts. This CRISPR/Cas9-mediated gene disruption tool is critical for exploring oxidative stress responses and hydrogen peroxide metabolism.
HEK293T cells, derived from human embryonic kidney and transformed with adenovirus 5 DNA, stably express SV40 large T antigen. This epithelial-like cell line is renowned for high transfectability, viral packaging, and recombinant protein production, making it an ideal host for knockout models. The robust proliferative capacity and genetic tractability of HEK293T provide a controlled environment for studying oxidative stress biology and downstream signaling pathways following CAT disruption.
Catalase is an essential antioxidant enzyme that decomposes hydrogen peroxide into water and oxygen, shielding cells from oxidative damage. Its transcription is activated by FOXO3 and NFE2L2 (Nrf2), with additional regulation by PPARG, TP53, and NF-??B. Functionally, catalase interacts with superoxide dismutase 1 (SOD1), glutathione peroxidase, and peroxiredoxins to detoxify reactive oxygen species. Downstream of the NFE2L2?CKEAP1 axis, catalase reduces H2O2 levels, thereby modulating MAPK signaling, NF-??B activity, and suppressing caspase-3/9-mediated apoptosis to maintain redox homeostasis.
CRISPR/Cas9-mediated knockout of CAT in HEK293T polyclonal cells abolishes catalase activity, causing intracellular hydrogen peroxide buildup and elevated basal oxidative stress. This sensitizes the cells to oxidative insults, enabling precise study of redox-sensitive pathways. In the context of actively proliferating HEK293T cells, the model reveals how catalase deficiency influences NF-??B and MAPK pathways. It thus provides a powerful system to explore the impact of oxidative stress on proliferation, survival, and metabolic adaptation.
These polyclonal knockout cells are ideal for antioxidant screening via DCFDA flow cytometry, drug toxicity assessment under H2O2 stress, and catalase validation by RT-qPCR or Western blotting. They also support research in aging, cancer biology, and metabolic disorders where catalase dysfunction is key. For further inquiries, please contact Ascent Research.
