ACOX1 Knockout HEK293T Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal population with targeted disruption of the human ACOX1 gene. This knockout product enables loss-of-function studies in a heterogeneous pool of HEK293T cells, eliminating the need for single-cell cloning while ensuring reliable gene inactivation. The polyclonal format is ideally suited for experiments requiring reproducible knockout effects and downstream phenotypic analyses in a well-characterized human cell background.
The host cell line, HEK293T, originates from human embryonic kidney epithelial cells and stably expresses the SV40 large T-antigen, conferring high transfection efficiency and robust growth. These adherent cells are widely used for genetic manipulation and functional assays due to their epithelial physiology and compatibility with standard culture protocols. Their active peroxisomal machinery and lipid metabolism pathways make them an appropriate model for investigating peroxisomal enzyme functions, including ACOX1-mediated fatty acid beta-oxidation.
ACOX1 encodes the first and rate-limiting enzyme of peroxisomal fatty acid beta-oxidation, catalyzing the desaturation of acyl-CoAs to 2-trans-enoyl-CoAs. This step is transcriptionally regulated by PPAR??, which responds to fatty acid ligands and peroxisome proliferators. Downstream metabolism of enoyl-CoA by enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, and 3-ketoacyl-CoA thiolase generates acetyl-CoA and shortened acyl-CoA chains. Within the peroxisome, ACOX1 interacts with PEX5 and other components of the multi-enzyme beta-oxidation complex. Disruption of ACOX1 therefore blocks peroxisomal VLCFA degradation, causing substrate accumulation and potentially activating PPAR?? signaling, which alters cellular energy homeostasis and lipid metabolism.
In HEK293T cells, ACOX1 knockout models key aspects of peroxisomal acyl-CoA oxidase deficiency and related Zellweger spectrum disorders. The epithelial context allows investigation of how VLCFA accumulation triggers oxidative stress, impacts mitochondrial function, and modulates PPAR??-dependent gene expression. These cells serve as a tractable system to dissect pathogenic mechanisms underlying pseudo-neonatal adrenoleukodystrophy and to evaluate interventions that restore peroxisomal function or bypass the metabolic block.
These knockout cells are applicable to diverse functional studies, including fatty acid oxidation assays, lipidomics profiling, and PPAR signaling analysis. Researchers can validate ACOX1 disruption via western blotting or RT-qPCR, assess metabolic flux, monitor peroxisomal morphology by immunofluorescence, and measure VLCFA levels. Additional assays such as ROS detection and cell viability challenges under fatty acid loading enable comprehensive phenotypic characterization. For further information, please contact Ascent Research.
