The CAT Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated from the A-549 human lung adenocarcinoma epithelial cell line by targeted disruption of the CAT gene. This loss-of-function model enables researchers to investigate catalase-dependent processes without the confounding variables of clonal selection. The polyclonal nature provides a heterogeneous population that retains the biological variability inherent to the parental line while abolishing catalase expression, offering a robust system for studying oxidative stress responses in a cancer-relevant context.
The parental A-549 cell line, derived from the lung adenocarcinoma of a 58-year-old Caucasian male, is an extensively characterized model for lung carcinoma research. As an adherent epithelial cell line, A-549 recapitulates key features of alveolar type II pneumocytes and is widely employed in studies of epithelial barrier function, drug transport, and oncogenic signaling. Its inherent susceptibility to oxidative stress and well-documented responsiveness to reactive oxygen species (ROS) make it particularly suitable for dissecting the contributions of antioxidant enzymes such as catalase.
Catalase, encoded by CAT, is a peroxisomal antioxidant enzyme that catalyzes the decomposition of hydrogen peroxide into water and molecular oxygen, thereby protecting cells from oxidative damage. Catalase expression is transcriptionally regulated by several key factors, including FOXO3a, NRF2 (NFE2L2), and PPAR??, and its activity is modulated by upstream signals such as TNF-?? and insulin. Within the antioxidant network, catalase acts downstream of superoxide dismutase 1 (SOD1) and interacts with peroxins (PEX5, PEX14) for proper peroxisomal import. This enzyme works in concert with other components of the ROS detoxification system, including glutathione peroxidase 1 (GPX1) and the NRF2-KEAP1 axis, to maintain cellular redox homeostasis. Catalase-mediated scavenging of hydrogen peroxide prevents oxidative DNA damage and preserves the integrity of cellular proteins and lipids.
In A-549 cells, knockout of CAT is expected to disrupt the cellular capacity to detoxify hydrogen peroxide, leading to an accumulation of ROS and consequent oxidative stress. This model recapitulates aspects of acatalasemia and other oxidative stress-related disorders, providing a platform to examine how loss of catalase function sensitizes lung cancer cells to oxidative insults. Given that A-549 cells harbor oncogenic mutations and exhibit altered redox metabolism, the CAT knockout allows researchers to probe the interplay between antioxidant defense, ROS-mediated signaling, and malignant phenotypes, including proliferation, apoptosis, and drug resistance.
The CAT Knockout A-549 Polyclonal Cells are suited for a diverse range of experimental applications, including oxidative stress studies, cancer biology research, and drug response screening under conditions of impaired hydrogen peroxide metabolism. Typical downstream assays include Western blotting and RT-qPCR for verifying catalase ablation, DCFDA-based ROS detection, direct quantification of hydrogen peroxide levels, and viability assays following H?O? challenge. Furthermore, the cells facilitate Comet assay-based assessment of oxidative DNA damage and apoptosis assays to elucidate cell death pathways. This product is a valuable tool for antioxidant research and redox signaling investigations. For additional technical information or custom inquiries, please contact Ascent Research.
