The CAT knockout NCI-H1299 polyclonal cells are a CRISPR/Cas9-edited mixed cell population derived from the NCI-H1299 human non-small cell lung cancer line, featuring targeted disruption of the catalase (CAT) gene. This polyclonal knockout format provides a heterogeneous pool of loss-of-function variants, generated by CRISPR/Cas9-mediated gene disruption, enabling robust population-level analysis of catalase deficiency without clonal selection or isolation. The product offers a versatile tool for investigating the cellular consequences of impaired hydrogen peroxide metabolism in a disease-relevant pulmonary adenocarcinoma background.
The NCI-H1299 host cell line is a p53-deficient metastatic lung adenocarcinoma model isolated from a lymph node metastasis. These adherent cells exhibit epithelial morphology and are widely employed in cancer research, drug screening, and oncogenic signaling studies due to their characteristic genetic alterations and aggressive phenotype. The p53-null background sensitizes cells to oxidative stress and alters DNA damage response pathways, making the line particularly informative for dissecting interactions between tumor suppressor loss and redox homeostasis. Combined with catalase knockout, this model accentuates vulnerability to reactive oxygen species (ROS)-induced damage and aberrant survival signaling.
Catalase encoded by CAT is a peroxisomal heme enzyme that catalyzes the decomposition of hydrogen peroxide (H2O2) into water and molecular oxygen, serving as a primary antioxidant defense. At the molecular level, catalase expression is transcriptionally regulated by forkhead box O3a (FOXO3a), nuclear factor erythroid 2-related factor 2 (NRF2), and peroxisome proliferator-activated receptor gamma (PPAR??), and is induced by oxidative stress, hypoxia, and cytokines including tumor necrosis factor alpha (TNF-??) and interleukin-1?? (IL-1??). Catalase functionally interacts with superoxide dismutase (SOD) and glutathione peroxidase (GPX) while its import into peroxisomes relies on the peroxisomal import receptor PEX5. Loss of catalase disrupts H2O2 detoxification, leading to elevated intracellular ROS that can activate downstream redox-sensitive targets such as nuclear factor-kappa B (NF-??B), activator protein-1 (AP-1), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK), ultimately affecting cell survival, proliferation, and metabolic adaptation.
In the NCI-H1299 lung cancer context, catalase knockout creates a model for studying how antioxidant failure exacerbates oncogenic phenotypes. The absence of functional p53, a known regulator of ROS metabolism and ferroptosis, combined with catalase deficiency, promotes a pro-oxidative state that may alter the activity of HIF-1?? signaling, enhance mutagenic pressure, and modify sensitivity to chemotherapeutic agents. This system permits researchers to dissect the interplay between tumor suppressor loss and oxidative stress management, including the contributions of the NRF2/KEAP1 axis and glutathione (GSH) systems in sustaining viability. The model is thus valuable for probing mechanisms of redox adaptation, drug resistance, and metabolic reprogramming in non-small cell lung cancer.
Research applications of the CAT knockout NCI-H1299 polyclonal cells span oxidative stress biology, cancer redox signaling, and therapeutic response profiling. The population can be used in assays including Western blotting and RT-qPCR for catalase and pathway validation, catalase activity measurement, DCFDA-based ROS detection, H2O2 quantification, MTT/XTT viability tests, caspase-3 activation apoptosis assays, wound healing migration studies, and chemosensitivity screening with cisplatin or doxorubicin. Additionally, immunofluorescence for peroxisomal localization and analysis of interacting partners like SOD and GPX support mechanistic investigations into antioxidant network disruption. For further details or technical support, please contact Ascent Research.
