The FAHD2A Knockout Raji Polyclonal Cells product offers a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji human B lymphocyte cell line, in which the FAHD2A gene has been disrupted to generate a loss-of-function model. This polyclonal pool contains a heterogeneous mixture of edited cells, enabling the study of functional consequences of FAHD2A ablation without isolating single-cell clones. By disrupting the target gene through CRISPR/Cas9-mediated gene editing, this model facilitates investigations into FAHD2A’s role in mitochondrial metabolism and cancer biology.
The Raji cell line is an EBV-positive Burkitt’s lymphoma line exhibiting a lymphoblastoid phenotype and retaining features of mature B lymphocytes. Widely used in immunological and oncology research, Raji cells provide a relevant model for studying lymphomagenesis, adaptive immunity, and the metabolic reprogramming that accompanies malignant transformation. Their high proliferative rate and reliance on aerobic glycolysis and glutaminolysis make them particularly suitable for examining mitochondrial functions and metabolic dependencies in cancer.
FAHD2A encodes a mitochondrial oxaloacetate decarboxylase that converts oxaloacetate to pyruvate, thus connecting oxaloacetate metabolism with pyruvate pools and NAD+ regeneration. This enzyme is transcriptionally regulated by MYC and metabolic stress, and functions downstream of mitochondrial biogenesis regulators PGC-1?? and NRF1. FAHD2A activity influences key metabolic intermediates including pyruvate, NAD+, and acetyl-CoA, and its interaction with oxaloacetate in the mitochondrial matrix is functionally linked to malate dehydrogenase and the TCA cycle. Disruption of FAHD2A decarboxylase activity alters oxaloacetate levels, impairs TCA cycle flux, and compromises NAD+ homeostasis, thereby reducing metabolic plasticity.
In the context of Raji Burkitt’s lymphoma cells, which exhibit heightened MYC-driven metabolic demands, FAHD2A knockout is expected to exacerbate metabolic vulnerabilities by limiting the cell??s ability to modulate pyruvate and oxaloacetate levels. This disruption may sensitize the lymphoma cells to metabolic stress, as the TCA cycle becomes less adaptable and NAD+ regeneration is compromised. Consequently, this model serves as a valuable tool for probing the metabolic checkpoints that govern survival and proliferation in aggressive B-cell malignancies, and for identifying nodes that may be exploited therapeutically.
Researchers can employ this polyclonal knockout model in a range of experimental workflows to dissect mitochondrial metabolism and cancer cell adaptation. Representative applications include metabolic vulnerability screening using Seahorse metabolic flux analysis, LC-MS metabolomics to profile TCA cycle intermediates, and NAD+/NADH assays to quantify redox balance. The cells are also suitable for cell proliferation and apoptosis assays, as well as RT-qPCR and western blotting to examine compensatory changes in metabolic gene expression. For further details or technical support, please contact Ascent Research.
