The HADHA Knockout HeLa Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated by disrupting the HADHA gene in HeLa cells, yielding a loss-of-function model for the mitochondrial trifunctional protein alpha subunit. This polyclonal pool ensures robust target-gene disruption and is ideal for functional genomics and metabolic studies without clonal selection artifacts.
The host, HeLa, is a human cervical adenocarcinoma epithelial cell line immortalized by HPV18 and characterized by p53 inactivation. Widely employed in cancer and cell biology, HeLa provides a well-defined background for exploring HADHA-dependent metabolic pathways, given its rapid proliferation and tractable metabolism.
HADHA encodes the alpha subunit of the mitochondrial trifunctional protein, which together with HADHB forms an inner mitochondrial membrane complex catalyzing the final steps of long-chain fatty acid beta-oxidation: hydration, dehydrogenation, and thiolysis. Upstream, PPARA, PPARGC1A, SIRT1, and insulin/glucagon signaling regulate expression and activity, while electron transfer proteins ETFA and ETFB, and chaperone HSPD1, facilitate proper function. Downstream, HADHA activity generates acetyl-CoA, NADH, FADH2, and ATP, and modulates ROS. Disruption thus halts fatty acid-derived energy production and elevates oxidative stress.
HADHA knockout in HeLa cells recapitulates mitochondrial trifunctional protein deficiency, producing accumulation of long-chain acylcarnitines and fatty acids, impaired mitochondrial respiration, and increased ROS. This model is particularly valuable in cancer metabolism research, as many tumors rely on fatty acid oxidation for energy and biosynthetic substrates; HADHA loss may unveil metabolic vulnerabilities and synthetic lethal interactions.
Key applications include modeling long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency and general mitochondrial trifunctional protein deficiency, as well as screening compounds to ameliorate fatty acid oxidation disorders. Cancer-focused studies can leverage this model to dissect lipid metabolic reprogramming, using assays such as [3H]-palmitate oxidation flux, acylcarnitine profiling, Seahorse respirometry, ATP and ROS quantification, BODIPY lipid staining, TOMM20 immunofluorescence, and viability under metabolic stress. For additional product information, contact Ascent Research.
