ECHS1 Knockout HeLa Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population targeting the ECHS1 gene in the HeLa cell line. This loss-of-function model enables study of mitochondrial fatty acid ??-oxidation and related metabolic pathways. The polyclonal pool contains a heterogeneous mix of edited cells, minimizing clonal artifacts and maintaining genetic diversity.
HeLa cells are an immortalized human cervical adenocarcinoma cell line derived from a 31-year-old African American woman. They contain integrated human papillomavirus 18 (HPV18) sequences, contributing to continuous proliferation. HeLa cells are widely used in cancer biology, drug metabolism, and metabolic studies due to their robust growth and high transfection efficiency. Their epithelial origin and metabolic flexibility make them suitable for investigating mitochondrial function and lipid metabolism.
ECHS1 encodes short-chain enoyl-CoA hydratase, a mitochondrial enzyme that catalyzes the second step of fatty acid ??-oxidation: the hydration of trans-2-enoyl-CoA to 3-hydroxyacyl-CoA. This reaction is critical for the breakdown of short-chain fatty acids and branched-chain amino acids (valine, leucine, isoleucine). ECHS1 is transcriptionally regulated by PPAR??, PGC-1??, and HNF4??, and its activity produces acetyl-CoA and NADH for the TCA cycle and electron transport chain. The enzyme interacts with short-chain acyl-CoA dehydrogenase (SCAD) and electron transfer flavoprotein (ETF) and cooperates with HADH, ACADS, and HADHA to complete fatty acid oxidation. Disruption of ECHS1 leads to accumulation of enoyl-CoA intermediates and impaired energy production.
In HeLa cells, ECHS1 knockout disrupts mitochondrial fatty acid oxidation, potentially shifting metabolic reliance toward glucose. This model recapitulates aspects of metabolic stress observed in ECHS1 deficiency disorders such as Leigh syndrome and recurrent metabolic decompensation. The polyclonal knockout pool in a cancer background allows investigation of metabolic reprogramming. Because HeLa cells express functional ??-oxidation machinery, ECHS1 loss-of-function provides a platform to study compensatory metabolic rewiring and the role of fatty acid oxidation in sustaining cancer cell proliferation under nutrient-limited conditions.
Researchers can use this model for fatty acid oxidation flux assays with labeled palmitate, mitochondrial respiration measurements by Seahorse analysis, and acylcarnitine profiling via metabolomics. It is suitable for investigating ECHS1 deficiency pathogenesis, screening small-molecule modulators of fatty acid metabolism, and examining cellular stress responses. Western blotting and RT-qPCR confirm gene disruption and assess compensatory enzyme expression. This product supports academic and pharmaceutical research on mitochondrial disorders and metabolic vulnerabilities in cancer. For additional information, please contact Ascent Research.
