The CAT Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population for functional analysis of catalase, a critical antioxidant enzyme encoded by the CAT gene. This polyclonal pool contains a heterogeneous mix of HAP1 cells carrying diverse loss-of-function mutations at the target locus, enabling robust population-level studies without clonal bias. Designed for direct use in oxidative stress pathway dissection, this knockout model eliminates catalase activity, allowing researchers to examine the consequences of impaired hydrogen peroxide (H2O2) detoxification in a defined genetic background.
The host HAP1 cell line is a human near-haploid chronic myeloid leukemia-derived model, originating from the male KBM-7 line. Its near-haploid karyotype simplifies genetic knockout studies by removing the complexity of diploid gene redundancy, making HAP1 cells a widely adopted platform for functional genomics, drug target validation, and pooled CRISPR screening. The haploid nature ensures that a single disruptive mutation can produce a complete loss of protein function, offering a clear readout in downstream assays.
Catalase is a peroxisomal heme-containing enzyme that decomposes H2O2 into water and oxygen, serving as a primary defense against reactive oxygen species (ROS). Transcriptionally regulated by FOXO3, NFE2L2 (NRF2), and PPARG, and activated by AKT1 and MAPK1/3 signaling, CAT interacts with peroxisomal import receptors PEX5 and PEX14 for proper subcellular localization. It functions in concert with superoxide dismutase 1 (SOD1) and glutathione peroxidase 1 (GPX1) to form a coordinated antioxidant network, where SOD1 generates H2O2 from superoxide, and both CAT and GPX1 independently detoxify H2O2. CAT specifically prevents H2O2-induced protein carbonylation, lipid peroxidation, and DNA damage, and its activity is linked to cellular senescence and stress response pathways.
Disruption of CAT in HAP1 cells results in elevated intracellular ROS levels, heightened sensitivity to oxidative challenges, and accumulation of oxidative damage markers, including ??H2AX foci and senescence-associated ??-galactosidase staining. This model is particularly valuable for dissecting peroxisomal antioxidant defense mechanisms and for evaluating the role of catalase in pathologies such as acatalasemia, diabetes, hypertension, and Alzheimer’s disease. The polyclonal format reduces off-target concerns by averaging effects across diverse mutations, making it suitable for high-throughput chemical or genetic modifier screens.
Researchers can employ these cells in a range of assays including catalase activity measurements, DCFDA-based flow cytometry for ROS quantification, Western blotting, qRT-PCR, and immunofluorescence for peroxisomal markers. They are ideal for oxidative stress pathway analysis, antioxidant efficacy testing, chemotherapy sensitization studies, and as controls in pooled CRISPR screens. For further technical details or to request a quote, please contact Ascent Research.
