The ECHDC3 Knockout HeLa Polyclonal Cells consist of a genetically heterogeneous HeLa cell population generated through CRISPR/Cas9-mediated disruption of the ECHDC3 gene. This polyclonal knockout model serves as a powerful tool to investigate the functional role of mitochondrial enoyl-CoA hydratase in cellular metabolism. The pooled format avoids the clonal selection artifacts often associated with single-cell-derived lines, ensuring that experimental observations reflect the average behavior of a diverse knockout population.
HeLa cells are a human cervical adenocarcinoma epithelial line that harbors HPV-18 sequences and exhibits an aggressive proliferative phenotype. Extensively characterized across decades of research, these cells provide a reliable and consistent system for studying oncogenic signaling, metabolic reprogramming, and mitochondrial function. Their robust growth and well-annotated genome make HeLa cells an ideal host for generating knockout models, particularly for genes involved in lipid metabolism and energy homeostasis.
ECHDC3 encodes a mitochondrial enoyl-CoA hydratase that catalyzes the second step of fatty acid beta-oxidation, hydrating trans-2-enoyl-CoA to 3-hydroxyacyl-CoA. It is a core component of the mitochondrial fatty acid synthesis pathway, forming functional complexes with MECR, HSD17B8, OXSM, and ACSM3. ECHDC3 expression is transcriptionally regulated by PPARA and PPARG, key nuclear receptors that govern lipid catabolism, and is modulated by AMPK and insulin signaling in response to energy status. Downstream, ECHDC3 activity is linked to HADH, a mitochondrial dehydrogenase, and contributes to the synthesis of lipoic acid, an essential cofactor for TCA cycle enzymes. Disruption of ECHDC3 abrogates proper beta-oxidation flux, potentially leading to accumulation of medium-chain acyl-CoA species and metabolic imbalance.
In the HeLa cancer cell context, where mitochondrial function is adapted to support rapid proliferation, ECHDC3 knockout uncovers dependencies on fatty acid oxidation. The polyclonal knockout population enables systematic evaluation of how loss of this enzyme reprograms mitochondrial substrate utilization, impairing oxygen consumption and triggering compensatory glycolysis. This model is particularly valuable for exploring the intersection of beta-oxidation defects with cancer cell survival, especially under nutrient-limited or oxidative stress conditions that challenge mitochondrial integrity.
The ECHDC3 Knockout HeLa Polyclonal Cells are suitable for diverse assays, including targeted metabolic flux analysis with Seahorse analyzers, fatty acid oxidation rate quantification, and comprehensive lipidomic profiling to detect changes in acyl-carnitines and phospholipids. Protein and transcript levels of key nodes such as MECR, HADH, and OXSM can be assessed by Western blotting and RT-qPCR. Mitochondrial membrane potential and ATP output measurements further delineate the bioenergetic phenotype. This knockout resource is adaptable for high-throughput drug screens against metabolic targets in oncology and metabolic disease. For further details or to place an order, please contact Ascent Research.
