The GLS2 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the GLS2 gene has been disrupted in the human Burkitt lymphoma-derived Raji B lymphocyte cell line. This polyclonal knockout model offers a heterogeneous pool of loss-of-function alleles, enabling robust functional investigation of GLS2-dependent processes without clonal selection bias. The knockout cell population serves as an advanced tool for dissecting glutamine metabolism, redox regulation, and p53-associated signaling in a malignant B-cell background, supporting applications in cancer biology, immunology, and drug discovery.
The host cell line, Raji, is an EBV-positive, suspension-adapted B lymphocyte model originally isolated from a Burkitt lymphoma patient. Raji cells exhibit key features of aggressive B-cell lymphoma, including rapid proliferation and constitutive survival signaling, making them a widely used system for studying lymphomagenesis, immune cell interactions, and therapeutic responses. Their B lymphoblastoid origin preserves relevant oncogenic and metabolic pathways, providing a physiologically relevant context for interrogating gene functions that impact malignant transformation and lymphocyte biology.
GLS2 encodes the mitochondrial glutaminase 2 enzyme, which catalyzes the hydrolysis of glutamine to glutamate, fueling the TCA cycle through ??-ketoglutarate production and contributing to glutathione (GSH) biosynthesis. Transcriptionally activated by TP53 (p53) and modulated by regulators such as NRF2 (NFE2L2), mTORC1, HIF1A, and SIRT5, GLS2 sits at a critical node linking cellular energy metabolism, redox homeostasis, and ferroptosis sensitivity. It interacts with SIRT5, BNIP3, glutamate dehydrogenase (GLUD1), and HSP90, and its activity influences downstream targets including ATP, NADPH, mTORC1 activity, ROS levels, and caspase activation. This positions GLS2 as a key mediator of metabolic and stress signals that govern cell proliferation and apoptosis.
In the Raji lymphoma context, GLS2 disruption illuminates the intersection between glutamine dependency and p53-regulated ferroptosis. Lymphoma cells often exhibit heightened glutamine consumption, and GLS2??s role in supplying glutamate for antioxidant defense and anaplerosis is particularly relevant in Burkitt lymphoma, where MYC-driven metabolic demands and p53 pathway alterations are common. By eliminating GLS2 function in this model, researchers can probe how the loss of p53-inducible glutaminase activity alters metabolic flux, sensitizes cells to oxidative damage, and reshapes survival signaling, revealing vulnerabilities that may be exploited therapeutically.
Typical applications encompass glutamine metabolism profiling via glutaminase activity assays and glutamine/glutamate quantification, p53 pathway dissection with Western blotting and RT-qPCR, ferroptosis assessment using C11-BODIPY and glutathione measurements, and anti-cancer drug screening, particularly for glutaminase inhibitors. Complementary techniques such as Seahorse metabolic flux analysis, flow cytometry for proliferation and apoptosis (Annexin V/PI), ROS detection, and co-immunoprecipitation for protein interactions further expand the model’s utility. This GLS2 knockout population thus supports detailed investigation of metabolic and signaling networks in B-cell lymphoma. For further information or customized inquiries, please contact Ascent Research.
