The DPH5 Knockout Jurkat Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout population of the Jurkat T-lymphocyte cell line, carrying targeted disruption of the DPH5 gene. The polyclonal nature ensures representation of diverse editing events, enhancing experimental robustness when assessing gene function. This population serves as a loss-of-function model for studying diphthamide biosynthesis, translation elongation, and bacterial toxin susceptibility.
Jurkat cells, an immortalized human T lymphocyte line derived from the peripheral blood of a patient with acute T cell leukemia, are a cornerstone in T cell signaling and leukemia research. Their transformed phenotype and active signaling pathways such as IL-2, TCR, and integrin cascades enable detailed investigation of T cell activation, apoptosis, and oncogenic processes.
DPH5 encodes a methyltransferase that catalyzes trimethylation of diphthine at histidine-715 of eEF2, the penultimate step in diphthamide biosynthesis. This modification is essential for translation elongation and is the target for ADP-ribosylation by diphtheria and Pseudomonas exotoxin A. DPH5 acts within a complex containing DPH1, DPH2, DPH3, DPH4, DPH6, and DPH7, using S-adenosyl methionine. Constitutively expressed DPH5 is indispensable for generating functional diphthamide on eEF2, impacting ribosomal translocation.
In Jurkat T cells, DPH5 disruption eliminates diphthamide modification, conferring resistance to diphtheria toxin- and exotoxin A-induced killing while potentially altering translation elongation dynamics. This knockout model allows dissection of how translation control intersects with T cell proliferation, survival, and leukemogenesis. Additionally, Jurkat sensitivity to ADP-ribosylating toxins makes this system ideal for studying toxin uptake and cell death mechanisms in a lymphoid context.
Researchers can employ this polyclonal knockout model in various experimental settings to elucidate molecular mechanisms. Applications include investigating diphthamide biosynthesis via Western blotting for diphthamide-modified eEF2, performing ADP-ribosylation assays using recombinant toxins, conducting cell viability assays with diphtheria toxin to quantify toxin susceptibility, and verifying gene disruption through RT-qPCR for DPH5 transcript levels and Sanger sequencing. The product is also suitable for translation elongation studies and genome-wide functional screens in T-cell leukemia. For further technical details, please contact Ascent Research.
