The NDUFAF7 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in the Raji B lymphocyte line, with targeted disruption of the NDUFAF7 gene. This polyclonal format comprises a heterogeneous mixture of edited cells, minimizing clonal selection bias and providing a robust loss-of-function model. The Raji line, an Epstein-Barr virus-positive Burkitt lymphoma, serves as a well-characterized and widely used platform for B cell biology and oncology research.
Derived from a Burkitt lymphoma patient, Raji cells express surface immunoglobulin and MHC class II molecules, recapitulating key aspects of antigen-presenting B cells. Their continuous proliferation is driven by EBV immortalization, facilitating scalable and reproducible in vitro assays. Raji cells are extensively employed to study B lymphocyte metabolism, including mitochondrial respiration and glycolytic flux, making them an ideal host for dissecting NDUFAF7-dependent metabolic pathways.
NDUFAF7 encodes a mitochondrial matrix methyltransferase that catalyzes the methylation of arginine-85 on the NDUFS2 subunit of complex I, an essential post-translational modification for complex I assembly and stability. Its expression is regulated by PGC-1??, NRF1, NRF2, and TFAM, placing it downstream of master mitochondrial biogenesis programs. Knockout disrupts NDUFS2 methylation, leading to defective complex I activity, reduced oxidative phosphorylation, ATP depletion, and elevated ROS. NDUFAF7 interacts with assembly factors ECSIT, ACAD9, TMEM126B, and NDUFAF8, and is functionally linked to pathway components including NDUFS1, NDUFV1, and other subunits of complex I.
In Raji B cells, NDUFAF7 knockout generates a model of mitochondrial complex I deficiency within a hematopoietic lineage. B lymphocytes undergo metabolic reprogramming during activation and differentiation; intact oxidative phosphorylation is increasingly recognized as critical for certain B cell subsets and for malignant B cell survival. Disruption of NDUFAF7 permits dissection of how complex I dysfunction impacts B cell proliferation, antibody production, and apoptotic signaling. The model also enables studies of the interplay between EBV-driven oncogenic signals and mitochondrial metabolism, as well as the sensitivity of lymphoma cells to metabolic stress. Elevated ROS and ATP depletion further provide a system for investigating redox balance.
These polyclonal knockout cells support a wide range of functional assays, including Seahorse-based oxygen consumption rate (OCR) measurement, direct complex I enzymatic activity assays, Western blotting for NDUFS2 methylation status, ATP luminescence, MitoSOX staining for mitochondrial ROS, and JC-1 mitochondrial membrane potential analysis. Flow cytometry enables simultaneous detection of B cell surface markers (e.g., CD19, CD20) and viability, facilitating high-throughput screening of compounds that modulate complex I activity. Applications include investigating metabolic reprogramming in immune cells, modeling mitochondrial complex I deficiency disorders such as Leigh syndrome, and evaluating targeted therapeutics. For further information or custom inquiries, please contact Ascent Research.
