The DLAT Knockout AGS Polyclonal Cells are a human CRISPR/Cas9-edited polyclonal knockout cell population, offering a loss-of-function model for the DLAT gene in the AGS gastric adenocarcinoma cell line. This product comprises a heterogeneous pool of AGS cells with targeted disruption of DLAT, which encodes the dihydrolipoamide S-acetyltransferase (E2) subunit of the pyruvate dehydrogenase (PDH) complex. The polyclonal knockout enables bulk functional studies without clonal bias.
AGS is a widely used human gastric adenocarcinoma cell line with epithelial morphology, derived from a primary tumor. It serves as a model for studying gastric cancer biology, drug response, and metabolic regulation. This background is particularly suited for investigating genes governing central carbon metabolism, as AGS cells exhibit metabolic pathways relevant to tumorigenesis.
DLAT catalyzes acetyl transfer from S-acetyldihydrolipoamide to CoA, producing acetyl-CoA??a critical step linking glycolysis to the TCA cycle. DLAT functions in a multienzyme complex with PDHA1, PDHB, DLD, and PDHX, requiring lipoic acid. Its activity is controlled by the NADH/NAD+ ratio and by phosphorylation via PDKs and PDPs. Downstream products include acetyl-CoA, NADH, citrate, and ATP. DLAT is also regulated by HIF-1?? under hypoxia and by PGC-1?? during mitochondrial biogenesis.
In AGS gastric cancer cells, DLAT loss disrupts pyruvate-derived acetyl-CoA synthesis, altering energy metabolism and biosynthetic pathways. This model is valuable for studying metabolic reprogramming in cancer, including the effects of hypoxia and nutrient stress. The knockout facilitates examination of how DLAT deficiency impacts cell proliferation, migration, and survival, and can be paired with pharmacological inhibition of PDKs or TCA cycle enzymes to explore therapeutic strategies.
Researchers can assess PDH complex activity enzymatically, measure metabolic fluxes with Seahorse analyzers, and confirm DLAT disruption by Western blotting or RT-qPCR. Functional consequences, such as changes in proliferation and migration, are readily quantified in the AGS background. These cells are suitable for drug screens targeting the PDH complex or downstream metabolism. For further details, contact Ascent Research.
