NUDT15 Knockout Jurkat E6.1 Cell Line

The NUDT15 Knockout Jurkat E6.1 Cell Line is a human CRISPR/Cas9-edited knockout cell line originating from the Jurkat E6.1 T lymphoblast line. It carries a targeted disruption of the NUDT15 gene, resulting in a stable loss-of-function model for exploring nucleotide pool sanitization and thiopurine drug responses. The suspension-adapted Jurkat E6.1 host, derived from a 14-year-old acute T-cell leukemia patient, provides a well-characterized platform for T-cell signaling and leukemia research.

Jurkat E6.1 cells are a widely used human T-cell leukemia line that recapitulates key features of T-cell receptor (TCR) signaling and acute lymphoblastic leukemia (ALL). These cells exhibit immature T-lymphoblast morphology and respond robustly to TCR pathway stimuli, making them an established model for dissecting signal transduction. Their facile genetic manipulation and suspension growth facilitate high-throughput drug screening and CRISPR-based editing. In the NUDT15 knockout background, the fundamental TCR signaling and leukemogenic properties are retained, allowing specific interrogation of nucleotide metabolism pathways.

NUDT15 is a nucleoside triphosphate hydrolase that converts mutagenic nucleotides such as 8-oxo-dGTP and 6-thio-dGTP to monophosphates, blocking their DNA incorporation. Its expression is induced by the transcription factor NRF2 during oxidative stress, creating a protective link between redox status and nucleotide pool integrity. The enzyme associates with PCNA at replication foci, coupling sanitization to DNA synthesis. Loss of NUDT15 leads to accumulation of oxidized nucleotides, misincorporation by DNA polymerases, and consequent ??H2AX foci formation, driving apoptosis upon thiopurine challenge. Compensatory base excision repair (BER) activity amplifies replication stress and genomic instability.

Within the Jurkat E6.1 T-ALL context, the NUDT15 knockout affords a clean genetic background to dissect thiopurine sensitivity mechanisms. Thiopurines such as 6-mercaptopurine are cornerstone ALL therapies, yet NUDT15 variants are major determinants of dose-limiting myelosuppression. This isogenic model permits quantification of drug-induced apoptosis via Annexin V assays, measurement of DNA damage by ??H2AX immunofluorescence, and nucleotide pool analysis by LC-MS. It supports pharmacogenomic validation and screening for modulators of thiopurine toxicity, free from confounding patient heterogeneity.

Key applications include thiopurine toxicity profiling, DNA damage response studies, leukemia pharmacogenomics, and nucleotide metabolism research. Typical assays are Western blot and RT-qPCR for NUDT15 confirmation, 6-thioguanine sensitivity assays, and apoptosis detection. The cell line also serves in complementation experiments to evaluate clinical NUDT15 alleles. By providing a defined, loss-of-function system, the NUDT15 Knockout Jurkat E6.1 Cell Line accelerates translational research in cancer chemotherapy. For additional technical details, please contact Ascent Research.