The L2HGDH Knockout Jurkat Polyclonal Cells product comprises a polyclonal population of Jurkat T lymphocytes genetically engineered using CRISPR/Cas9 to disrupt the L2HGDH gene. This heterozygous population of edited cells provides a powerful loss-of-function tool for investigating the impact of L2HGDH deficiency on T cell biology without single-cell clonal isolation. The polyclonal format preserves cellular heterogeneity while ensuring robust gene disruption across the population, enabling functional studies in a biologically relevant context. Researchers can employ these cells to dissect the role of L2HGDH in metabolic and epigenetic regulation within an immortalized T cell background.
Jurkat cells are an extensively characterized human T lymphocyte line derived from the peripheral blood of a 14-year-old male patient with acute T cell leukemia. These immortalized cells serve as a widely adopted model for studying T cell receptor (TCR) signaling, apoptosis, and HIV infection. Their rapid proliferation and well-defined signaling networks make them an ideal host for generating gene knockout models. By introducing L2HGDH disruption in this well-established leukemic background, the resulting cell population offers a tractable system for examining the intersection of mitochondrial metabolism, oncometabolite accumulation, and T cell malignancy.
L2HGDH encodes a mitochondrial enzyme that catalyzes the oxidation of L-2-hydroxyglutarate (L-2HG) to alpha-ketoglutarate (??-KG), thereby preventing the accumulation of the oncometabolite L-2HG. This reaction relies on FAD as a cofactor and is transcriptionally regulated by TFAM and NRF1. Loss of L2HGDH function leads to elevated L-2HG levels, which competitively inhibit ??-KG-dependent dioxygenases, including TET DNA demethylases and JmjC-domain histone demethylases. Consequently, downstream epigenetic processes such as DNA hydroxymethylation and histone methylation are disrupted. This mechanistic link positions L2HGDH at a critical node connecting mitochondrial metabolism to chromatin regulation and gene expression.
In the Jurkat T cell leukemia context, L2HGDH knockout recapitulates key aspects of L-2-hydroxyglutaric aciduria and oncometabolite-driven malignancies. The resulting L-2HG accumulation is expected to impair epigenetic homeostasis, potentially altering the expression of genes involved in T cell activation, differentiation, and apoptosis. This model is particularly relevant for probing how metabolic dysregulation contributes to leukemogenesis and for evaluating the role of TET and JmjC enzymes in lymphoid cancers. Moreover, it enables the investigation of ??-KG-dependent prolyl hydroxylases (PHDs) that regulate hypoxia-inducible factor stability, adding a dimension of oxygen-sensing pathway analysis.
Key experimental applications include quantifying L2HGDH mRNA and protein levels via RT-qPCR and Western blotting to confirm gene disruption, and measuring L-2HG accumulation through LC-MS to validate the metabolic consequence. Chromatin immunoprecipitation?CqPCR (ChIP-qPCR) for histone methylation marks and assessment of global DNA hydroxymethylation can reveal epigenetic changes. Flow cytometry-based assays permit analysis of T cell activation markers, proliferation, and apoptosis in the knockout background. These cells are well-suited for genetic screens, inhibitor studies targeting residual dioxygenase activity, and rescue experiments. For additional information or to inquire about this model, please contact Ascent Research.
