IDH1 Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts IDH1 gene expression in a heterogeneous pool of Jurkat cells. This polyclonal format provides a population-level loss-of-function model without clonal isolation, and gene disruption is achieved through CRISPR/Cas9-mediated targeting, resulting in ablation of functional IDH1 protein across the edited pool. The product is intended for studies requiring a representative IDH1-deficient background in a T-lymphocyte context.
The Jurkat host cell line is a human CD4+ T-lymphocyte leukemia line derived from an acute lymphoblastic leukemia patient. These suspension cells are extensively used to investigate T-cell receptor signaling, cytokine production, and immune responses. Their robust growth, well-defined signaling networks, and relevance to T-cell biology and leukemia make them an ideal chassis for engineering targeted knockouts that probe metabolic and oncogenic mechanisms.
IDH1 encodes cytosolic isocitrate dehydrogenase 1, a homodimeric enzyme that converts isocitrate to ??-ketoglutarate (??-KG) while generating NADPH. Knockout of IDH1 eliminates this activity, decreasing ??-KG and NADPH. Consequently, ??-KG-dependent dioxygenases??prolyl hydroxylases targeting HIF-1??, TET DNA demethylases, and Jumonji histone demethylases??are inhibited, stabilizing HIF-1?? and disrupting DNA and histone methylation. Key downstream effects involve glutathione reductase and HIF-1?? transcriptional targets. IDH1 is regulated by c-Myc, SREBP1, AMPK, and stressors, and interacts with citrate synthase and aconitase.
In Jurkat T-cells, IDH1 loss disrupts redox homeostasis by limiting NADPH supply for antioxidant defense, potentially sensitizing cells to oxidative damage. The diminished ??-KG availability compromises TET-mediated DNA demethylation and Jumonji-dependent histone demethylation, which can rewire gene expression programs critical for T-cell activation, proliferation, and leukemic transformation. Additionally, HIF-1?? stabilization may promote a glycolytic shift, modeling metabolic reprogramming observed in cancers. Thus, this knockout model enables dissection of how IDH1 coordinates metabolic intermediates, redox status, and epigenetic regulation in immune cells.
Applications include metabolic flux analysis, oxidative stress assessment, drug sensitivity screens against IDH1-mutant cancers, and epigenetic studies. Recommended assays: Western blot, RT-qPCR, LC-MS for ??-KG, 2-hydroxyglutarate, and NADPH/NADP+, flow cytometry for apoptosis and proliferation, ROS detection, DNA methylation sequencing, and ChIP-qPCR for histone marks. For further technical inquiries, please contact Ascent Research.
