The DLAT Knockout 786-O Polyclonal Cells are a CRISPR/Cas9-edited heterogeneous population of 786-O cells with targeted disruption of the DLAT gene. This polyclonal knockout model provides a pooled loss-of-function system, eliminating clonal selection bias and enabling population-level studies of dihydrolipoamide S-acetyltransferase (E2) deficiency in a cancer cell context.
The parental 786-O line is derived from a clear cell renal cell carcinoma (ccRCC) with biallelic VHL inactivation, leading to constitutive HIF-1?? stabilization. This drives a pseudohypoxic metabolic phenotype marked by elevated glycolysis and suppressed mitochondrial respiration, establishing 786-O cells as a key model for cancer metabolism research and metabolic adaptation in ccRCC.
DLAT encodes the E2 subunit of the pyruvate dehydrogenase (PDH) complex, which catalyzes acetyl group transfer to coenzyme A, linking glycolysis to the TCA cycle. The PDH complex comprises E1 (PDHA1/PDHB), E2 (DLAT), E3 (DLD), and PDHX, and is regulated by PDK1?C4 (inactivating kinases) and PDP1/2 (activating phosphatases). HIF-1?? upregulates PDK1, inhibiting PDH and promoting glycolytic metabolism. DLAT interacts with PDHA1, PDHB, DLD, and PDHX; its loss disrupts complex formation, impairing acetyl-CoA synthesis and downstream oxidative phosphorylation, citrate production, and protein acetylation.
In 786-O cells, VHL deficiency and HIF-1?? activity already suppress PDH function, and DLAT ablation further cripples this metabolic node. The resulting shift toward aerobic glycolysis creates a robust system for examining PDH complex dependency in ccRCC, identifying metabolic vulnerabilities, and evaluating potential synthetic lethal targets under pseudohypoxic conditions.
These cells support metabolic flux assays (Seahorse), PDH subunit expression analysis (western blot, RT-qPCR), and metabolite quantification (acetyl-CoA, glucose, lactate). Functional studies on proliferation, migration, invasion, and apoptosis under metabolic stress, combined with RNA-seq profiling, can elucidate molecular consequences of DLAT loss. Applications include drug screening for PDH deficiency and HIF-1?? pathway interrogation. For technical information, contact Ascent Research.
