The ACOXL Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human ACOXL gene in HEK293T cells. This polyclonal pool consists of a heterogeneous mixture of cells carrying loss-of-function mutations, providing a robust loss-of-function model for examining peroxisomal fatty acid beta-oxidation. The CRISPR/Cas9-mediated gene disruption avoids clonal selection, enabling high-throughput functional screens and biochemical analyses while maintaining biological variability representative of complex metabolic states.
HEK293T cells are a widely used human embryonic kidney cell line that stably expresses the SV40 large T antigen, facilitating episomal plasmid replication and high-level protein expression. These adherent cells display epithelial morphology and are highly transfectable, making them an ideal host for gene-editing applications. Their metabolic capacity and ease of culture support a broad range of biochemical and functional studies, including those focused on lipid metabolism and peroxisomal biology.
The ACOXL protein is an acyl-CoA oxidase-like enzyme that catalyzes the initial desaturation step in peroxisomal fatty acid beta-oxidation, converting acyl-CoAs to 2-trans-enoyl-CoAs. Its expression is regulated by the nuclear receptor PPAR?? in response to long-chain fatty acids, placing ACOXL downstream of lipid sensing pathways. The enzyme operates upstream of a peroxisomal multienzyme complex that includes enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase, which complete the beta-oxidation cycle. ACOXL likely interacts with ACOX1 and other peroxisomal oxidases to coordinate acyl-CoA flux. Disruption of ACOXL activity therefore impairs the desaturation reaction, leading to accumulation of very long-chain fatty acids, altered lipid homeostasis, and downstream metabolic stress.
In HEK293T cells, peroxisomal beta-oxidation contributes to cellular lipid balance, particularly under conditions of high fatty acid supply or metabolic challenge. The ACOXL knockout polyclonal population enables dissection of ACOXL-specific functions without clonal bias, capturing the range of phenotypic responses to gene disruption. This model allows investigation of compensatory metabolic adaptations, crosstalk with mitochondrial oxidation, and the impact on membrane lipid composition, providing a physiologically relevant system for exploring peroxisomal disorder mechanisms.
This knockout cell product is suited for a variety of research applications, including the study of peroxisomal lipid metabolism and metabolic disease modeling. Researchers can measure very long-chain fatty acid accumulation by GC-MS, assess fatty acid oxidation rates with radiolabeled substrates, and characterize gene expression changes by RT-qPCR and western blotting. Lipidomics and cell viability assays under fatty acid challenge further define the metabolic stress phenotype. For inquiries regarding custom gene editing or additional cell models, please contact Ascent Research.
