The NT5C Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the gene encoding cytosolic 5′-nucleotidase III (NT5C) is disrupted. This loss-of-function model is generated in the Raji host cell background to enable investigation of nucleotide salvage pathway dependencies and responses to nucleoside analog chemotherapies. The polyclonal format preserves heterogeneous editing events, avoiding clonal selection artifacts and providing a population-level representation of NT5C ablation. Researchers can employ these cells for metabolomic, signaling, and drug-response studies without the constraints of a single clonal genotype.
The Raji cell line, derived from an EBV-positive Burkitt lymphoma patient, serves as a well-established model for B-cell biology, lymphomagenesis, and immune response studies. Characterized by CD20 positivity and the absence of surface immunoglobulin, Raji cells recapitulate key features of aggressive B-cell lymphomas. Their EBV association further positions them as a relevant system for studying viral contributions to oncogenesis. This background makes Raji a physiologically relevant host to examine how nucleoside metabolism impacts lymphoma viability and drug sensitivity.
NT5C catalyzes dephosphorylation of nucleoside monophosphates (AMP, CMP, UMP, dAMP, dCMP, dTMP) to nucleosides, serving as a key enzyme in purine and pyrimidine salvage. Its activity is regulated by substrate availability and the AMP/ATP ratio, with nucleoside analog drugs acting as competitive substrates. Downstream, NT5C impacts adenosine signaling, AMPK activity, and DNA synthesis by modulating nucleoside/nucleotide pools. It interacts with ENT1, CNT transporters, adenosine kinase, and deoxycytidine kinase, which together govern nucleoside uptake and phosphorylation. Within this network, NT5C functions alongside adenosine deaminase, purine nucleoside phosphorylase, hypoxanthine-guanine phosphoribosyltransferase, uridine-cytidine kinase, and thymidine kinase 1.
Disruption of NT5C in Raji lymphoma cells perturbs salvage-driven nucleotide homeostasis, potentially increasing dependence on de novo synthesis or alternative salvage pathways. This metabolic vulnerability is particularly relevant given the high proliferative demand of lymphomas. The knockout may enhance sensitivity to nucleoside analogs such as cytarabine and gemcitabine by reducing their inactivation through dephosphorylation, providing a model to study chemoresistance mechanisms. Additionally, altered adenosine production can affect AMPK-mediated energy sensing and modulate apoptotic thresholds in these EBV-positive B cells.
The NT5C Knockout Raji Polyclonal Cells support diverse research applications, including dose-response drug sensitivity assays with nucleoside analogs (cytarabine, gemcitabine), LC-MS-based nucleotide pool profiling, and functional assays for apoptosis (Annexin V, caspase activation) or viability (MTS, CellTiter-Glo). Western blot analysis for NT5C and phosphorylated AMPK, along with RT-qPCR for ENT1 and CNT transporters, facilitates mechanistic correlation. The polyclonal format is particularly suited for synthetic lethality screens seeking metabolic vulnerabilities in aggressive B-cell lymphomas. For further technical details and ordering information, please contact Ascent Research.
