The HSD17B8 knockout Jurkat polyclonal cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population derived from the immortalized human T lymphocyte Jurkat cell line. This product is designed for researchers studying mitochondrial fatty acid synthesis (mtFAS) and its role in T-cell metabolism and leukemia. The polyclonal nature of the knockout ensures a heterogeneous population with targeted disruption of the HSD17B8 gene, facilitating robust functional genomics studies without clonal selection artifacts. By ablating HSD17B8 expression, this model enables loss-of-function investigations into a critical enzyme of the mtFAS pathway, providing a physiologically relevant system to dissect metabolic dependencies in immune cells. The knockout cells are generated using CRISPR/Cas9 technology to deliver stable gene disruption, and they are supplied as a mixed population suitable for downstream biochemical, metabolic, and phenotypic analyses.
The Jurkat host cell line is a widely employed model in immunology and cancer research, originally established from the peripheral blood of a 14-year-old boy with acute T-cell leukemia. These cells exhibit features of mature T lymphocytes and have been extensively used to study T-cell receptor signaling, apoptosis, and leukemogenesis. Their robust growth characteristics, genetic tractability, and well-characterized signaling networks make them an ideal platform for examining metabolic pathways in leukemic T cells. Jurkat cells retain many aspects of primary T-cell biology while offering the experimental advantages of an immortalized line, including high transfection efficiency and reproducibility. This background provides a relevant cellular context for exploring how dysregulation of mitochondrial metabolism contributes to malignant transformation and immune cell dysfunction.
HSD17B8 encodes a mitochondrial 3-ketoacyl-ACP reductase that catalyzes the reduction of 3-ketoacyl-ACP to 3-hydroxyacyl-ACP within the mtFAS pathway, a process essential for the synthesis of lipoic acid and maintenance of mitochondrial lipid composition. The enzyme functions as part of a multienzyme complex alongside NDUFAB1 (acyl carrier protein), OXSM (3-ketoacyl-ACP synthase), MECR (trans-2-enoyl-ACP reductase), and MCAT (malonyl-CoA-ACP transacylase). HSD17B8 activity is transcriptionally regulated by upstream factors including PPARGC1A, NRF1, ESRRA, and PPARD, which integrate metabolic and proliferative signals. Downstream, its catalytic product 3-hydroxyacyl-ACP feeds into subsequent mtFAS steps, ultimately contributing to the biosynthesis of octanoylated mitochondrial proteins and lipoic acid, a critical cofactor for ??-ketoacid dehydrogenase complexes. Disruption of HSD17B8 therefore impairs mtFAS flux, leading to altered mitochondrial lipid homeostasis and compromised lipoic acid-dependent metabolic pathways.
In the Jurkat T-cell context, HSD17B8 knockout provides a powerful tool for dissecting the intersection of mitochondrial lipid metabolism and immune cell function. T lymphocytes undergo dramatic metabolic reprogramming upon activation, and emerging evidence implicates mtFAS in supporting proliferation and effector responses. The loss of HSD17B8 perturbs mitochondrial lipid remodeling necessary for oxidative phosphorylation and membrane integrity, which may sensitize leukemic cells to metabolic stress. This model enables mechanistic studies of how mtFAS influences T-cell signaling, apoptosis resistance, and metabolic plasticity. Moreover, because Jurkat cells are derived from a T-cell leukemia, the HSD17B8 knockout helps identify metabolic vulnerabilities that could be exploited therapeutically in hematological malignancies. The polyclonal knockout population, with its diverse mutational spectrum, mimics the heterogeneity of tumor cell populations, enhancing translational relevance.
Researchers can apply the HSD17B8 knockout Jurkat polyclonal cells in a variety of experimental workflows to investigate mitochondrial metabolism and its impact on T-cell biology. Western blotting and RT-qPCR confirm target gene disruption, while mitochondrial respiration assays using Seahorse analyzers assess oxidative phosphorylation and glycolysis. Fatty acid oxidation assays, lipidomics profiling, and analysis of lipoic acid content quantify metabolic consequences of HSD17B8 loss. Cell proliferation and apoptosis assays further evaluate functional outcomes, linking mtFAS to cell viability and growth. These applications support studies in metabolic disorders, cancer biology, and immunometabolism, facilitating identification of novel regulatory nodes and therapeutic targets. For additional information or custom services, please contact Ascent Research.
