The CAT Knockout HT29 Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the catalase (CAT) gene in the human HT29 colorectal adenocarcinoma cell line. This product provides a reliable loss-of-function model for investigating catalase-dependent hydrogen peroxide detoxification and its broader role in cellular redox homeostasis. The polyclonal pool ensures a heterogeneous knockout background, offering a robust system for functional studies without the genetic biases inherent to single-cell-derived clones. Applications span from basic oxidative stress biology to translational cancer research, where catalase deficiency is linked to altered proliferation, apoptosis, and tumorigenic potential.
HT29 cells are an established epithelial colorectal adenocarcinoma line widely used in cancer biology, drug development, and studies of intestinal epithelium. Originating from a primary tumor, they retain many characteristics of colorectal cancer, including oncogenic mutations and dysregulated signaling pathways. Their adherent growth and suitability for diverse assays make them an ideal platform for interrogating gene function in a therapeutically relevant context. When combined with CAT knockout, this model enables precise dissection of oxidative stress responses in a colorectal cancer setting, facilitating investigations into tumor microenvironment adaptation and resistance mechanisms.
Catalase is a peroxisomal enzyme that catalyzes the decomposition of hydrogen peroxide to water and oxygen, protecting cells from oxidative damage to lipids, proteins, and DNA. Its expression is transcriptionally regulated by factors such as PPAR??, Nrf2, and FoxO proteins, which respond to varying levels of oxidative stress. In the knockout model, disrupted catalase activity leads to accumulation of intracellular hydrogen peroxide, altering redox-sensitive signaling cascades. Catalase functions in concert with superoxide dismutase and glutathione peroxidase, and its absence shifts the balance toward oxidative injury, impacting pathways like FoxO signaling and longevity regulation. This signaling network positions catalase as a central node in cellular oxidant detoxification.
The introduction of CAT knockout into HT29 cells creates a physiologically relevant model to study oxidative stress in colorectal cancer. As colonocytes are frequently exposed to reactive oxygen species from microbial metabolism and inflammation, impaired hydrogen peroxide clearance may exacerbate DNA damage and promote genomic instability, hallmarks of colorectal carcinogenesis. The polyclonal population allows observation of heterogeneous cellular responses, such as variable susceptibility to apoptosis or proliferation changes under oxidative challenge. This model can be utilized to screen compounds that target redox vulnerabilities in cancer cells, evaluate the interplay between catalase and oncogenic signaling, and explore mechanisms of resistance to pro-oxidant therapies.
Typical applications include quantifying reactive oxygen species using DCFDA probes, measuring catalase activity and protein levels by Western blotting, assessing cell viability via MTT assays under oxidative stress, and analyzing apoptosis with Annexin V staining. Transcriptomic profiling by RNA-seq and DNA damage evaluation with comet assays further elucidate downstream consequences of catalase deficiency. This knockout pool serves as a versatile tool for dissecting antioxidant defense mechanisms and ROS-mediated signaling in colorectal adenocarcinoma research. For comprehensive technical data, custom cell engineering projects, or to request a quotation, please contact Ascent Research.
