The ACADVL Knockout HeLa Polyclonal Cells consist of a heterogeneous HeLa population in which the ACADVL gene has been disrupted by CRISPR/Cas9-mediated editing. This polyclonal knockout pool offers a non-clonal loss-of-function model for studying very long-chain acyl-CoA dehydrogenase (VLCAD) function, avoiding the phenotypic fixation of individual clones. The cell population is well-suited for metabolic assays that benefit from genetic diversity, such as population-level flux analyses and drug response profiling.
HeLa cells, derived from a cervical adenocarcinoma, are an immortalized epithelial line that is HPV18-positive and aneuploid. Widely used as a cancer cell model, they provide a resilient background for genetic perturbation and subsequent metabolic phenotyping. This epithelial context is particularly relevant for investigating the interplay between oncogenic transformation and mitochondrial fatty acid oxidation, as well as for translational studies in cervical and other epithelial cancers.
ACADVL encodes VLCAD, a mitochondrial enzyme that initiates beta-oxidation of long-chain fatty acids (C14?CC20) by catalyzing their ??,??-dehydrogenation, utilizing FAD as a cofactor. The reaction feeds electrons via electron transfer flavoprotein (ETF) to the respiratory chain. ACADVL expression is promoted by PPAR?? and PGC-1?? in response to fasting and glucagon, and its activity generates acetyl-CoA, ATP, and long-chain acylcarnitines. Disruption thus impairs fatty acid-derived energy production and alters lipid metabolic signaling.
Within the HeLa model, ACADVL knockout reveals metabolic vulnerabilities that arise from defective long-chain fatty acid oxidation. Cancer cells frequently rewire metabolism to support proliferation, and loss of VLCAD can unmask dependencies on alternative substrates or pathways. This model enables the dissection of how mitochondrial dysfunction influences redox balance, ATP supply, and lipid intermediate accumulation, providing insight into metabolic adaptations in HPV-driven cervical adenocarcinoma and beyond.
These polyclonal knockout cells are amenable to a range of assays, including RT-qPCR and immunoblotting for confirmation of ACADVL disruption, 3H-palmitate oxidation to measure beta-oxidation flux, acylcarnitine profiling by LC-MS, and Seahorse analysis of oxygen consumption. Applications include drug screening for very long-chain acyl-CoA dehydrogenase deficiency (VLCADD), studies of metabolic cardiomyopathy and myopathy, and PPAR?? agonist testing. For further details or customized services, please contact Ascent Research.
