The DLAT Knockout NCI-H1975 Polyclonal Cells product comprises a population of CRISPR/Cas9-edited polyclonal knockout cells derived from the NCI-H1975 non-small cell lung adenocarcinoma cell line. This loss-of-function model is generated through CRISPR/Cas9-mediated gene disruption of DLAT, the gene encoding dihydrolipoamide S-acetyltransferase (E2 subunit of the pyruvate dehydrogenase complex). The polyclonal format provides a heterogeneous pool of alleles, offering a versatile tool for studying metabolic rewiring without the selection bottlenecks associated with clonal cell lines.
The NCI-H1975 cell line, derived from the pleural effusion of a 62-year-old female with non-small cell lung adenocarcinoma, is a widely used model retaining oncogenic drivers and metabolic dependencies of lung adenocarcinoma. Its epithelial adherent properties suit monolayer and 3D culture for studies in cancer biology and metabolic adaptation.
DLAT encodes the E2 subunit of the pyruvate dehydrogenase complex (PDC), catalyzing acetyl group transfer from pyruvate to CoA to generate acetyl-CoA and link glycolysis to the TCA cycle. Its activity is regulated by PDK1-mediated phosphorylation and PDP1-mediated dephosphorylation, integrating pyruvate, insulin, and NADH/NAD+ signals. Acetyl-CoA downstream fuels lipid synthesis via ACLY and ACC1 and histone acetylation via HATs. DLAT functions in complex with PDHA1, PDHB, DLD, PDHX, and lipoic acid cofactor. Disruption severs this metabolic conduit.
CRISPR/Cas9-mediated knockout of DLAT in NCI-H1975 cells eliminates the E2 subunit, dismantling PDC activity and abolishing mitochondrial pyruvate-to-acetyl-CoA conversion. The polyclonal knockout cells undergo a forced metabolic shift to aerobic glycolysis, mimicking the Warburg effect with elevated lactate production and reduced TCA cycle flux, while impacting lipogenesis and histone acetylation. This model is valuable for studying metabolic reprogramming in lung adenocarcinoma and compensatory mechanisms that bypass DLAT.
This DLAT knockout model is well-suited for western blotting to confirm target protein loss and pyruvate dehydrogenase activity assays to validate functional knockout. It enables metabolic flux analyses via Seahorse XF, glucose uptake and lactate secretion assays, and metabolomic profiling of acetyl-CoA and TCA cycle intermediates. The polyclonal population can be employed in testing glycolytic inhibitors, PDK1 modulators, or drugs targeting ACLY, as well as in proliferation, clonogenic, and drug sensitivity assays. It also facilitates studies on metabolism-epigenetics crosstalk through histone acetylation measurements. For further inquiries, please contact Ascent Research.
