MDH1 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population originating from Raji human B lymphocytes. This product provides a mixed pool of edited cells with disrupted MDH1 alleles, offering a loss-of-function model to investigate the consequences of cytoplasmic malate dehydrogenase deficiency without clonal isolation.
The Raji host cell line was established from a Burkitt lymphoma patient and is Epstein?CBarr virus (EBV)-positive, exhibiting hallmark B-cell characteristics and robust proliferation. As a widely used model in immunology and oncology, Raji cells enable the study of B-lymphocyte biology, antibody production, and lymphomagenesis. The MDH1 knockout in this background allows dissection of metabolic liabilities specific to aggressive B-cell malignancies.
MDH1 encodes cytoplasmic malate dehydrogenase, which catalyzes the reversible oxidation of malate to oxaloacetate using NAD+ as a cofactor, a reaction that is fundamental to the malate?Caspartate shuttle and cytosolic NADH reoxidation. MDH1 expression is controlled by transcription factors HIF1A, MYC, and PPARGC1A, and is modulated by glucose availability and insulin signaling. The enzyme??s activity directly impacts oxaloacetate pools, the NAD+/NADH ratio, aspartate biosynthesis, gluconeogenesis, and lipogenesis. MDH1 functionally interacts with its mitochondrial counterpart MDH2, as well as with citrate synthase, malic enzyme, glutamate dehydrogenase, and aspartate aminotransferase, forming a metabolic hub that integrates carbohydrate, amino acid, and energy metabolism.
In Raji Burkitt lymphoma cells, MDH1 loss abrogates the malate?Caspartate shuttle, preventing the transfer of reducing equivalents from the cytosol to the mitochondrial matrix. This leads to a perturbed NAD+/NADH balance and diminished oxaloacetate availability, which can compromise biosynthetic pathways and redox homeostasis. Given the high glycolytic activity and MYC-mediated upregulation of MDH1 in these cells, the knockout may sensitize them to metabolic stress, highlighting potential therapeutic vulnerabilities.
Applications include metabolic reprogramming studies in lymphoma, NAD+/NADH redox biology, drug sensitivity screening, and lentiviral CRISPR knockout modeling. Researchers can characterize the model using Western blotting and RT-qPCR for MDH1, quantify NAD+/NADH ratios, measure metabolic flux via Seahorse analyzers, assess cell viability with MTT/XTT, monitor apoptosis with Annexin V, and profile cell cycle by flow cytometry. Downstream transcriptomic analysis by RNA-seq permits global assessment of MDH1-dependent gene networks. For further information, contact Ascent Research.
