DLAT Knockout Huh-7 Polyclonal Cells represent a genetically engineered cell population in which the DLAT gene has been disrupted via CRISPR/Cas9-mediated editing in the Huh-7 human hepatocellular carcinoma cell line. This polyclonal format consists of a pool of edited cells with varied targeting events, offering a robust and biologically relevant model for loss-of-function studies without the selection bias of clonal isolates. The product is designed for researchers investigating pyruvate dehydrogenase complex function, metabolic reprogramming, and oncogenic signaling in liver cancer.
The Huh-7 cell line, derived from a well-differentiated hepatocellular carcinoma of a 57-year-old Japanese male, is a widely employed model in liver biology and oncology. These cells maintain key hepatic functions, including metabolic and detoxification pathways, and exhibit robust glycolytic and oxidative metabolism. Their relevance to human liver cancer makes them an appropriate host for dissecting the impact of DLAT ablation on cancer cell metabolism, mitochondrial activity, and tumorigenic properties.
DLAT encodes the dihydrolipoamide S-acetyltransferase (E2) component of the pyruvate dehydrogenase complex (PDC), which catalyzes the oxidative decarboxylation of pyruvate to acetyl?CoA, linking glycolysis to the TCA cycle. Within the PDC, DLAT interacts with PDHA1 (E1) and DLD (E3), and is functionally regulated by PDK isoforms such as PDK1 that phosphorylate E1, as well as the phosphatase PDP1. Upstream, DLAT expression is controlled by metabolic sensors PPARGC1A and HIF1A, and is responsive to insulin signaling. Downstream, acetyl-CoA generated by PDC feeds the TCA cycle, supports histone acetylation, and provides precursors for lipid synthesis. CRISPR-mediated DLAT disruption in Huh-7 cells therefore impedes pyruvate decarboxylation, leading to diminished acetyl-CoA and TCA cycle flux, impaired oxidative phosphorylation, and a compensatory increase in glycolytic dependence??mirroring metabolic reprogramming observed in aggressive cancers.
In hepatocellular carcinoma, dysregulation of the pyruvate dehydrogenase complex contributes to metabolic flexibility and tumor survival. The DLAT knockout Huh-7 model recapitulates features of pyruvate dehydrogenase deficiency and cancer metabolic rewiring, including reduced mitochondrial respiration and elevated lactate production. This system enables precise interrogation of how DLAT loss influences hepatocellular carcinoma cell proliferation, migration, and invasion, and allows for the identification of synthetic lethal interactions or metabolic liabilities that may be targeted with small-molecule inhibitors. Moreover, it provides a valuable platform for studying the consequences of PDH dysfunction on redox balance, epigenetic regulation via acetyl-CoA availability, and lipid metabolism.
These polyclonal knockout cells are amenable to a broad array of experimental analyses, including pyruvate dehydrogenase activity assays, Seahorse extracellular flux analysis to measure oxygen consumption and extracellular acidification, Western blotting for PDC components, RT?qPCR for metabolic gene expression, and LC?MS?based metabolomics. Key research applications encompass cancer metabolism studies, investigation of mitochondrial disorders, metabolic reprogramming in liver cancer, drug screening for PDH deficiencies, and functional validation of signaling nodes such as PDK1 and HIF1A. For further details on purchase, validation, and technical support, please contact Ascent Research.
