DLAT Knockout Raji Polyclonal Cells constitute a polyclonal knockout cell population generated by CRISPR/Cas9-mediated gene disruption of the DLAT gene in the human Raji B-lymphocyte cell line. This product enables loss-of-function studies of dihydrolipoamide S-acetyltransferase, the E2 component of the pyruvate dehydrogenase complex (PDC). The polyclonal format preserves heterogeneous editing outcomes, providing a robust model for investigating metabolic reprogramming in B-cell lymphoma.
The Raji host cell line is an Epstein-Barr virus-immortalized B lymphocyte derived from a Burkitt lymphoma patient. These cells express B-cell markers including CD19 and CD20, maintain a high proliferation rate, and grow in suspension, making them suitable for metabolic studies. As an EBV-positive lymphoma model, Raji cells are widely used to investigate oncogenic signaling, immune interactions, and metabolic adaptations in B-cell malignancies.
DLAT encodes the dihydrolipoamide S-acetyltransferase (E2) subunit of the pyruvate dehydrogenase complex (PDC), which catalyzes the transfer of an acetyl group from acetyl-TPP to lipoamide. Within the PDC, DLAT interacts with E1 subunits (PDHA1 and PDHB), the E3 component (DLD), and the E3-binding protein (PDHX). Its activity is tightly regulated by PDK family kinases (PDK1?C4), which phosphorylate and inhibit the E1?? subunit, and PDP phosphatases (PDP1?C2), which reverse this inhibition. Upstream signals including insulin, PPAR, and PGC-1?? influence PDC activity, while downstream outputs include generation of acetyl-CoA for the TCA cycle, histone acetylation mediated by acetyl-CoA availability, lipid biosynthesis, and NADH production. The mitochondrial deacetylase SIRT3 has been shown to interact with and modulate PDC components.
In Raji B-lymphoma cells, DLAT knockout disrupts the conversion of pyruvate to acetyl-CoA, impairing mitochondrial oxidative metabolism and promoting a metabolic shift toward glycolysis and glutaminolysis. This metabolic reprogramming mimics the Warburg effect commonly observed in aggressive lymphomas and reduces the availability of acetyl-CoA for histone acetylation and lipid biosynthesis. Consequently, this knockout model serves as a powerful tool to study how pyruvate dehydrogenase deficiency influences lymphoma cell proliferation, survival, and epigenetic regulation, as well as the adaptive mechanisms that B-cell malignancies employ under metabolic stress.
Researchers can use DLAT Knockout Raji Polyclonal Cells to investigate metabolic vulnerabilities in B-cell lymphoma through a variety of assays, including Western blot validation of DLAT and histone acetylation status, PDH enzymatic activity measurements, Seahorse extracellular flux analysis to assess oxygen consumption (OCR) and extracellular acidification (ECAR), 13C-pyruvate metabolic tracing to map carbon flux, and acetyl-CoA quantification. Additional applications include mitochondrial membrane potential assessment via JC-1 staining, drug sensitivity profiling against metabolic inhibitors, and CRISPR-based synthetic lethality screens. This model is particularly suited for dissecting the role of PDC in central carbon metabolism and evaluating therapeutic strategies targeting metabolic dependencies in lymphoma. For further details, please contact Ascent Research.
