The EHHADH Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-mediated gene disruption model targeting the human EHHADH gene, produced as a polyclonal knockout cell population in the HeLa cervical adenocarcinoma cell line. This product is engineered to serve as a robust in vitro tool for investigating peroxisomal fatty acid beta-oxidation and related metabolic disorders. The polyclonal format provides a heterogeneous population of knockout cells, enabling studies of loss-of-function effects without clonal selection bias. Researchers can employ this model to interrogate how EHHADH disruption impacts lipid catabolism and peroxisomal homeostasis, utilizing a range of biochemical and cell-based assays.
HeLa cells are an immortalized human cervical cancer line derived from HPV18-positive adenocarcinoma. Their biological context features inactivation of the tumor suppressors p53 and Rb by viral oncoproteins E6 and E7, resulting in deregulated cell cycle control and a metabolic profile favorable for examining cancer metabolic reprogramming. This background is commonly applied in cancer biology and drug screening, providing a well-characterized platform for evaluating genetic perturbations in pathways such as fatty acid degradation and PPAR signaling. The EHHADH knockout in HeLa cells allows dissection of peroxisomal function within a transformable and widely used host, facilitating reproducible experiments across lipid metabolism and oncology research.
EHHADH encodes a bifunctional enzyme harboring enoyl-CoA hydratase and 3-hydroxyacyl-CoA dehydrogenase activities, catalyzing the second and third reactions of peroxisomal fatty acid beta-oxidation. It hydrates trans-2-enoyl-CoA to 3-hydroxyacyl-CoA and subsequently dehydrogenates this intermediate to 3-ketoacyl-CoA, shortening very long-chain fatty acyl-CoA esters. EHHADH functions downstream of ACOX1 and upstream of ACAA1 within the peroxisomal beta-oxidation spiral, ultimately yielding acetyl-CoA and shorter-chain acyl-CoAs for mitochondrial oxidation or ketone body synthesis. Its activity is transcriptionally regulated by PPARA agonists such as fibrates, linking it to the PPAR signaling pathway. The protein requires interaction with PEX5 for peroxisomal import; disruption of EHHADH leads to accumulation of very long-chain fatty acids and perturbed lipid homeostasis.
In the HeLa background, EHHADH knockout provides a pertinent model for exploring peroxisomal dysfunction in the context of cancer metabolism. HPV18-driven transformation imposes unique lipid demands, and peroxisomal beta-oxidation may contribute to sustaining proliferation under nutrient stress. Loss of EHHADH can be exploited to investigate how impaired fatty acid degradation alters cellular energetics, mitochondrial crosstalk, and oncogenic signaling. Additionally, HeLa cells retain the core peroxisomal import machinery, making this model suitable for studying interactions with PEX5 and other peroxisomal enzymes. The knockout thereby bridges metabolic disorder research and cancer biology, offering insights into dual roles of peroxisomes in tumor cells.
This polyclonal knockout product is applicable to a spectrum of research areas, including peroxisomal biogenesis disorders, Zellweger spectrum disorders, and L-bifunctional enzyme deficiency modeling. It supports mechanistic studies of PPARA agonist action and downstream metabolic reprogramming, employing assays such as palmitate oxidation assays, peroxisomal beta-oxidation activity measurements, and very long-chain fatty acid profiling. The model also facilitates drug screening for modulators of peroxisomal metabolism and immunofluorescence-based analysis of peroxisomal markers. Broader applications extend to cancer lipidomics and metabolomics, examining acetyl-CoA flux and ketone body production. Researchers can integrate techniques like RT-qPCR and western blotting to validate target disruption and pathway readouts. For further details on applying the EHHADH Knockout HeLa Polyclonal Cells in your investigations, please contact Ascent Research.
