The HADH Knockout HeLa Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population targeting the HADH gene in the HeLa host cell background. This product provides a heterogeneous ensemble of loss-of-function variants for the hydroxyacyl-CoA dehydrogenase gene, enabling comprehensive interrogation of mitochondrial fatty acid ??-oxidation and insulin secretion regulatory pathways. The polyclonal format offers robustness against clonal variability, making it suitable for pathway dissection and pharmacological profiling in a well-established human cancer cell line.
HeLa cells, originally derived from the cervical adenocarcinoma of patient Henrietta Lacks, represent one of the most widely used epithelial cancer models in biomedical research. As an immortalized cervical epithelial line, HeLa cells exhibit rapid proliferation and hallmark metabolic adaptations characteristic of malignant transformation. Their robust and reproducible growth properties facilitate high-throughput genetic perturbation studies, particularly those exploring the intersection of oncogenic signaling and metabolic reprogramming.
HADH encodes the mitochondrial short-chain L-3-hydroxyacyl-CoA dehydrogenase, a pivotal enzyme in the ??-oxidation spiral. It catalyzes the NAD+-dependent oxidation of L-3-hydroxyacyl-CoA to 3-ketoacyl-CoA, transferring electrons to the electron transfer flavoprotein and generating NADH, which feeds into the respiratory chain for ATP synthesis. HADH activity is transcriptionally regulated by PPAR-?? and is responsive to the intracellular free fatty acid pool and the insulin/glucagon ratio. The enzyme functions within a multienzyme framework alongside mitochondrial trifunctional protein and interacts with NAD+ and electron transfer flavoprotein. In pancreatic ??-cells, HADH-mediated shifts in the NADH/NAD+ ratio influence membrane potential and insulin secretory dynamics, linking fatty acid catabolism to glucose homeostasis.
In the HeLa cervical adenocarcinoma context, HADH knockout disrupts a critical step in mitochondrial ??-oxidation, allowing direct assessment of metabolic vulnerability in cancer cells. This model is particularly relevant for dissecting the contribution of fatty acid oxidation to ATP generation, redox balance, and anaplerotic carbon flux in rapidly proliferating tumors. It also provides a platform to study the molecular underpinnings of familial hyperinsulinemic hypoglycemia type 3 and broader fatty acid oxidation disorders, facilitating the analysis of genotype-phenotype relationships without confounding backgrounds.
Researchers can employ this polyclonal knockout population in a range of functional assays, including NADH quantification, mitochondrial respiration profiling, and fatty acid oxidation rate measurements. Complementary techniques such as western blotting and RT-qPCR enable validation of downstream targets like 3-ketoacyl-CoA and acetyl-CoA pathway intermediates. In the context of metabolic syndrome, the model supports insulin secretion ELISA-based studies and apoptosis assays to investigate ??-cell-like metabolic coupling. Drug screening campaigns targeting metabolic modulators benefit from the population??s uniform genetic background while preserving allelic diversity. For additional information and ordering, please contact Ascent Research.
