The MTHFD1L Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, with targeted disruption of the MTHFD1L gene. This heterogeneous knockout pool lacks functional MTHFD1L protein, providing a loss-of-function model to study mitochondrial one-carbon metabolism without clonal selection.
The Raji parental cell line is an Epstein-Barr virus (EBV)-positive Burkitt lymphoma B lymphocyte line originally isolated from an 11-year-old male patient. Raji cells are extensively used in B-cell lymphoma research, exhibiting rapid proliferation and well-defined metabolic characteristics typical of MYC-driven tumors. Their EBV-positive status also makes them a relevant model for studying viral latency and oncogenic cooperation.
MTHFD1L encodes the mitochondrial NADP+-dependent 5,10-methylenetetrahydrofolate dehydrogenase/cyclohydrolase that oxidizes 5,10-methenyltetrahydrofolate to 10-formyltetrahydrofolate, producing NADPH and releasing formate. This reaction is a key step in mitochondrial one-carbon flux, coupling serine catabolism via SHMT2 and MTHFD2 to cytoplasmic purine nucleotide synthesis and mitochondrial translation initiation through formylation of methionyl-tRNA. MTHFD1L expression is transcriptionally activated by MYC and E2F factors and is responsive to folate availability, HIF1A, and metabolic stress. The enzyme functions within a mitochondrial one-carbon metabolism complex that includes SHMT2, MTHFD2, and ALDH1L2, channeling one-carbon units toward formate production and NADPH regeneration.
In the context of Raji B-cell lymphoma, knockout of MTHFD1L disrupts mitochondrial formate output, potentially impairing de novo purine biosynthesis and mitochondrial protein synthesis, which are critical for sustaining rapid proliferation. This model is valuable for dissecting MYC-driven metabolic dependencies in lymphomagenesis and for evaluating the metabolic consequences of folate pathway disruption in an EBV-positive background. The polyclonal population allows assessment of heterogeneous responses to metabolic challenges, reflecting the complexity of tumor cell populations.
Typical applications include metabolomic profiling by LC-MS to quantify folate intermediates, proliferation and clonogenic assays, flow cytometric analysis of cell cycle and apoptosis, and drug sensitivity testing with antifolates such as methotrexate and pemetrexed. Researchers can also employ this knockout model for CRISPR essentiality screens, mitochondrial translation assays, and functional complementation studies. For further information, custom services, or technical inquiries, please contact Ascent Research.
