The H6PD Knockout Jurkat Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the H6PD gene has been disrupted. This product offers a pooled population of Jurkat cells carrying heterogeneous mutations at the H6PD locus, enabling loss-of-function studies without clonal selection. CRISPR/Cas9-mediated gene disruption allows researchers to investigate the cellular consequences of H6PD ablation in a T-lymphoblastoid background.
The parental Jurkat cell line (clone E6-1) is an immortalized human T-lymphoblastoid line derived from an acute T-cell leukemia patient. Jurkat cells are widely used as a model for T-cell receptor signaling, apoptosis, and immunological studies due to their robust growth and well-characterized signaling pathways. Their malignant origin also makes them a relevant platform for oncology research, particularly in hematopoietic malignancies.
H6PD (hexose-6-phosphate dehydrogenase) localizes to the lumen of the endoplasmic reticulum (ER), where it catalyzes the conversion of glucose-6-phosphate (G6P) to 6-phosphogluconolactone, generating NADPH. This NADPH pool is essential for the activity of 11??-hydroxysteroid dehydrogenase type 1 (HSD11B1), which converts inactive cortisone to active cortisol. Thus, H6PD indirectly regulates glucocorticoid receptor signaling and downstream transcriptional programs. Upstream regulators include glucocorticoids, insulin, C/EBP transcription factors, and PPAR??, while downstream targets comprise HSD11B1, NADPH-dependent ER enzymes, and glucocorticoid-responsive genes. The pathway integrates metabolic cues (pentose phosphate pathway flux) with endocrine signaling (local cortisol regeneration), influencing adipocyte differentiation, insulin sensitivity, and cellular redox homeostasis.
In Jurkat T-lymphoblastoid cells, H6PD knockout provides a unique tool to dissect the intersection of glucocorticoid metabolism and T-cell biology. Given the well-established role of glucocorticoids in immune modulation and T-cell apoptosis, the loss of H6PD may alter cortisol-mediated responses, including gene expression changes and metabolic reprogramming. Moreover, Jurkat cells exhibit a high rate of glycolysis and pentose phosphate pathway activity, making them an apt model to study how ER NADPH production influences redox balance and biosynthetic capacity in malignant lymphocytes. This knockout therefore bridges immunology, endocrinology, and cancer metabolism.
Researchers can employ this knockout polyclonal population to investigate glucocorticoid metabolism in T cells via cortisone-to-cortisol conversion assays (HPLC/MS), assess ER redox status using NADPH/NADP+ ratio measurements, and monitor HSD11B1 expression by western blot. Metabolic flux analysis and RT-qPCR of glucocorticoid-responsive genes further enable detailed pathway dissection. Applications extend to studies of T-cell metabolism, metabolic syndrome, obesity, and apparent cortisone reductase deficiency. For further inquiries, please contact Ascent Research.
