The PFKP Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the human Raji B lymphocyte line, engineered for disruption of the PFKP gene. This heterogeneous pool of edited cells enables loss-of-function studies in a lymphoma-relevant background. The polyclonal format preserves cellular diversity without clonal isolation, making it suitable for population-level metabolic and functional analyses. The CRISPR/Cas9-mediated gene disruption ensures robust and stable PFKP knockout, providing a reliable model for examining the role of this glycolytic enzyme.
The Raji cell line originates from a Burkitt’s lymphoma patient and is a widely used model for B-cell malignancies. These cells are EBV-positive and harbor a t(8;14) translocation causing c-MYC overexpression, which drives rapid proliferation and a glycolytic metabolic phenotype. As B lymphocytes, Raji cells produce antibodies and present antigens, features retained in this knockout model. The EBV-driven transformation and c-MYC dysregulation make Raji cells highly dependent on glycolysis, mirroring the Warburg effect in aggressive lymphomas, thereby providing an ideal host to study PFKP-mediated metabolic vulnerabilities.
PFKP encodes the platelet-type phosphofructokinase, catalyzing the rate-limiting conversion of fructose-6-phosphate to fructose-1,6-bisphosphate in glycolysis. It is allosterically activated by AMP and inhibited by ATP and citrate, with expression transcriptionally controlled by MYC, HIF1A, and signaling through AMPK and mTOR. PFKP acts upstream of ALDOA, GAPDH, PKM, and LDHA, driving pyruvate and lactate production and ATP generation. PFKP also interacts with actin, linking glycolysis to the cytoskeleton. Disruption of PFKP attenuates glycolytic flux, reduces lactate output, and compromises energy homeostasis, particularly in glycolytic cancers.
In Raji cells, PFKP knockout is expected to cripple the Warburg effect, a hallmark of c-MYC- and EBV-driven malignancies. Loss of PFKP impairs the capacity for high glycolytic rates, leading to reduced ATP levels and metabolite accumulation, potentially inducing growth arrest or apoptosis. This model is invaluable for studying metabolic rewiring upon PFKP ablation, including possible compensatory activation of the pentose phosphate pathway or glutaminolysis. By examining PFKP dependency in this lymphoma context, researchers can evaluate its role in sustaining proliferation and survival under oncogenic stress imposed by MYC and EBV.
These polyclonal knockout cells support diverse applications, including glycolysis dependency studies in B-cell lymphomas, metabolic reprogramming analysis, Warburg effect characterization, anti-glycolytic target validation, and PFK inhibitor screening. Typical assays involve Western blotting, RT-qPCR, genomic sequencing, Seahorse metabolic flux analysis, lactate and ATP measurements, MTT proliferation assays, flow cytometry for apoptosis, and 2-deoxyglucose sensitivity testing. For further information, please contact Ascent Research.
