The NUDT1 Knockout Raji Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte cell line. This product introduces a targeted disruption of the human NUDT1 gene, which encodes the nucleotide sanitizing enzyme also known as MTH1. Designed as a heterogeneous pool of gene-edited cells, the model enables loss-of-function studies of NUDT1 within a lymphoblastoid background, providing a versatile tool for investigating oxidative nucleotide metabolism and DNA damage responses.
The Raji host cell line originates from a Burkitt lymphoma and maintains an Epstein-Barr virus (EBV)-positive status, growing in suspension as lymphoblastoid cells. Widely employed as a model for B cell malignancies and immune cell signaling, Raji cells exhibit characteristic features of high-grade B cell lymphoma, including dysregulated proliferation and susceptibility to DNA-damaging agents. This genetic background provides a clinically relevant platform for examining the interplay between nucleotide pool sanitization and lymphoid transformation.
NUDT1 functions as a critical housekeeping enzyme that hydrolyzes oxidized purine nucleoside triphosphates, most notably 8-oxo-dGTP and 2-hydroxy-dATP, into their corresponding monophosphates, thereby preventing their misincorporation into DNA and RNA. Its expression is transcriptionally regulated by the antioxidant transcription factor NRF2 (NFE2L2) in response to reactive oxygen species (ROS), with additional regulatory input from HIF1?? under hypoxia. Downstream of NUDT1 activity, 8-oxo-dGMP production leads to reduced intracellular 8-oxo-dGTP levels, mitigating the accumulation of 8-oxoguanine in DNA and diminishing the burden of oxidative DNA strand breaks and mutagenesis. This pathway intersects with base excision repair machinery, including DNA glycosylases OGG1 and MUTYH, which recognize and excise oxidized bases once incorporated.
Given that NUDT1 is frequently overexpressed in various cancers, including hematological malignancies, its disruption in the Raji lymphoma model is particularly informative. B cell lymphomas are often characterized by elevated replicative stress and altered redox homeostasis, conditions under which NUDT1-mediated nucleotide sanitization becomes essential for maintaining genomic integrity and cell viability. The loss of NUDT1 in this polyclonal population is expected to increase the abundance of oxidized nucleotides, potentially leading to elevated DNA damage and heightened sensitivity to oxidative insults, thereby allowing researchers to dissect the enzyme’s role in sustaining the malignant phenotype.
This polyclonal NUDT1 knockout product is suited for a spectrum of experimental applications, including functional dissection of oxidative DNA repair pathways, characterization of nucleotide pool sanitization mechanisms, and validation of small-molecule MTH1 inhibitors such as TH588 and (S)-crizotinib. Researchers can monitor NUDT1 expression by Western blotting and RT-qPCR, quantify 8-oxo-dGTPase activity through biochemical hydrolysis assays, and assess DNA damage accumulation via comet assays or ??H2AX immunofluorescence. Cellular responses to oxidative challenge can be evaluated by H2O2 treatment followed by viability, apoptosis, and cell cycle analyses using flow cytometry. Combining NUDT1 disruption with pharmacological inhibition enables comprehensive studies on synthetic lethality and therapeutic resistance in B cell malignancy models. For additional details, please contact Ascent Research.
