The NUBPL Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human Raji B-cell line, offering a loss-of-function model for the mitochondrial NUBPL gene. NUBPL encodes a critical iron-sulfur cluster transfer protein required for late-stage assembly of NADH:ubiquinone oxidoreductase (complex I). The polyclonal population contains diverse gene disruptions, enabling robust phenotypic analysis without clonal bias, and is suited for high-throughput studies of mitochondrial function in lymphoid cells.
The Raji host cell line is an Epstein-Barr virus (EBV)-positive B-lymphoblastoid line from Burkitt lymphoma, featuring suspension growth and expression of B-cell surface markers. Its transformed phenotype, driven by MYC and NF-??B activation and latent EBV genes, makes it a standard model for immunology and oncology. The suspension format facilitates metabolic flux assays and drug screening, and the lymphoma background provides a unique context for investigating mitochondrial dependencies.
NUBPL operates in mitochondrial iron-sulfur (Fe-S) cluster biogenesis, accepting clusters from the ISCU scaffold and inserting them into complex I subunits such as NDUFS1, NDUFS7, and NDUFV1. It functions within an assembly network including NDUFAF1, NFU1, BOLA3, HSCB, and HSPA9. Upstream transcription is driven by NRF1, PGC-1??, and TFAM, integrating with mitochondrial biogenesis programs. Disruption of NUBPL leads to complex I deficiency, reduced ATP synthesis, membrane potential collapse, and elevated reactive oxygen species.
In Raji B cells, NUBPL ablation exposes metabolic vulnerabilities critical to Burkitt lymphoma survival, which often relies on oxidative phosphorylation. This model enables discovery of synthetic lethal interactions and drug sensitivities under mitochondrial stress. Additionally, as NUBPL mutations cause Leigh syndrome and mitochondrial encephalopathy, the cells offer a unique lymphoid platform to study cell-type-specific complex I disease pathology, complementing conventional neuronal models.
Key applications include complex I enzyme activity assays, Western blot and blue-native PAGE for complex I analysis, Seahorse metabolic flux measurements, ATP quantification, and mitochondrial membrane potential assessments using JC-1 or TMRM. Further uses encompass ROS detection, viability testing under galactose challenge, and high-throughput drug screening targeting OXPHOS. This knockout tool is ideal for researching iron-sulfur cluster disorders in B-cells, metabolic adaptations in lymphoma, and mitochondrial disease therapeutic strategies. For further information, please contact Ascent Research.
