The HAGH Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of Jurkat T lymphocytes bearing a targeted disruption of the HAGH gene, encoding hydroxyacylglutathione hydrolase (glyoxalase II). This knockout product provides a genetically mixed model for studying loss of HAGH function in a human T-cell line, avoiding clonal selection artifacts while preserving the natural polyclonal diversity of edited cell pools.
Jurkat is an immortalized human T lymphocyte cell line derived from an acute T-cell leukemia patient, widely used to investigate T-cell signaling, activation, apoptosis, and adaptive immunity. The cells express CD3, CD4, and the T-cell receptor complex and proliferate robustly in suspension, making them amenable to gene editing and suitable for a variety of functional assays.
HAGH (glyoxalase II) catalyzes the hydrolysis of S-lactoylglutathione to D-lactate and glutathione in the glyoxalase pathway, a critical system for detoxifying methylglyoxal, a reactive glycolytic byproduct. HAGH expression is regulated by NFE2L2 (Nrf2) and other oxidative stress-responsive transcription factors, and its activity is essential for glutathione recycling and limiting advanced glycation end-product (AGE) formation. Knockout of HAGH disrupts methylglyoxal detoxification, leading to S-lactoylglutathione and methylglyoxal accumulation, increased glycation stress, and compromised redox balance. The glyoxalase pathway also includes GLO1, which produces S-lactoylglutathione, placing HAGH downstream of this reaction.
In the Jurkat T-cell background, HAGH knockout permits dissection of how carbonyl stress affects immune cell function and survival. T lymphocytes upregulate glycolysis upon activation, increasing methylglyoxal production; loss of glyoxalase II may impair T-cell receptor signaling, cytokine production, and viability. This model is also relevant to studying glyoxalase dysfunction in leukemia and diabetic complications, where elevated methylglyoxal contributes to pathogenesis.
Typical applications include glyoxalase II activity assays, methylglyoxal and glutathione quantification, western blotting for HAGH and AGEs, apoptosis assays, RT-qPCR for glyoxalase genes, and flow cytometry for ROS and cell viability. These polyclonal knockout cells enable investigation of the glyoxalase pathway in T-cell biology, methylglyoxal toxicity, and glutathione-dependent detoxification in hematopoietic cells. For further information or to order custom knockout cell products, please contact Ascent Research.
