The MTHFD2 Knockout Raji Polyclonal Cells are a polyclonal population of Raji B lymphocytes with CRISPR/Cas9-mediated disruption of the MTHFD2 gene. This engineered cell pool enables loss-of-function studies of mitochondrial one-carbon metabolism in a Burkitt lymphoma background. The polyclonal format captures diverse editing events, facilitating robust population-level phenotypic analysis without clonal bias. It is ideal for investigating MTHFD2-dependent metabolic processes supporting rapid lymphoma cell proliferation.
The Raji cell line is an EBV-positive lymphoblastoid line derived from a Burkitt lymphoma patient. As a B-lymphocyte model, it recapitulates key features of B-cell biology and lymphomagenesis, including high proliferative capacity and constitutive survival signaling. Raji cells are widely used to study oncogenic signaling, apoptosis, and metabolic reprogramming in hematological malignancies. The EBV-driven immortalization mimics metabolic demands of aggressive B-cell neoplasms, making Raji a relevant system to explore MTHFD2 function.
MTHFD2 is a mitochondrial bifunctional enzyme that converts methylene-THF to formyl-THF, providing one-carbon units for purine synthesis and generating NADPH. Its expression is induced by ATF4, c-MYC, HIF1A, and NRF2. MTHFD2 works intimately with SHMT2 and MTHFD1L within the mitochondrial folate cycle. Downstream, it sustains activities of TYMS, DHFR, and IMPDH, thereby maintaining nucleotide pools and redox balance. Disruption of MTHFD2 thus impairs de novo purine synthesis and diminishes NADPH, sensitizing cells to metabolic stress.
In Burkitt lymphoma, c-MYC-driven growth imposes high demand for one-carbon units and NADPH, rendering MTHFD2 a critical metabolic node. Raji cells with uncontrolled MYC activity rely on mitochondrial folate metabolism for biomass accumulation. Knocking out MTHFD2 in this context enables dissection of how mitochondrial folate cycle intersects with oncogenic signaling, potentially revealing synthetic vulnerabilities. The polyclonal knockout population is a valuable tool to probe metabolic dependencies in MYC-driven lymphomas and to evaluate therapeutic strategies targeting one-carbon metabolism.
Researchers can use this model for target disruption validation via Western blot and RT-qPCR, proliferation and colony formation assays, metabolic flux analysis, nucleotide pool measurements, and apoptosis assays. It is well-suited for drug target validation against antifolate agents and other one-carbon metabolism inhibitors, and for functional genomics screens identifying compensatory pathways. The polyclonal composition enhances reproducibility of population-level findings. For additional information, contact Ascent Research.
