The ACOXL Knockout HeLa Polyclonal Cells product consists of a heterogeneous pool of HeLa cells subjected to CRISPR/Cas9-mediated disruption of the ACOXL gene. This polyclonal knockout population provides a loss-of-function model for studying peroxisomal acyl-CoA oxidase function without clonal selection, enabling analysis of cellular responses across a genetically diverse background. The CRISPR-edited pool offers a practical tool for investigating ACOXL-dependent metabolic processes.
HeLa cells are an immortalized human cell line derived from a cervical adenocarcinoma, widely utilized as a model for epithelial cell biology and cancer research. Their robust proliferation, ease of culture, and well-characterized genetic and metabolic landscape make them suitable for investigating gene function in a tumorigenic context. These cells endogenously express machinery for peroxisomal lipid metabolism, providing a physiologically relevant platform to dissect the roles of peroxisomal enzymes like ACOXL.
ACOXL encodes a peroxisomal acyl-CoA oxidase that is predicted to catalyze the first step of fatty acid beta-oxidation. This enzyme functions downstream of the transcription factor PPAR-alpha, a master regulator of lipid catabolism. ACOXL activity is dependent on its import into the peroxisome via the PEX5 receptor and peroxisomal import machinery. Mechanistically, ACOXL interacts with the fatty acid oxidation multienzyme complex that includes enoyl-CoA hydratase and 3-ketoacyl-CoA thiolase. Disruption of ACOXL is expected to impair very long-chain fatty acid oxidation, leading to lipid droplet accumulation and altered cellular energy homeostasis.
In cervical adenocarcinoma cells, peroxisomal function is increasingly recognized for its contributions to tumor metabolism and stress adaptation. The ACOXL knockout in HeLa cells disrupts a critical node in peroxisomal lipid breakdown, potentially sensitizing cells to metabolic stress and exposing vulnerabilities in cancer cell bioenergetics. This model enables interrogation of how peroxisome-derived signals intersect with proliferative pathways in epithelial tumor cells, offering insights into the metabolic flexibility of cervical cancer.
Researchers can employ this polyclonal knockout model to examine roles of peroxisomal beta-oxidation in tumor cell proliferation, lipid metabolism, and drug sensitivity. Standard assays include immunoblotting for ACOXL to confirm protein loss, RT-qPCR for mRNA quantification, fatty acid oxidation rate measurements, peroxisome immunofluorescence staining, and global lipidomics profiling to map lipidome alterations. Additionally, the cells are suitable for screening small-molecule metabolic inhibitors that target fatty acid catabolism. For further information on assay customization or technical support, please contact Ascent Research.
