The ADPGK Knockout HEK293T Polyclonal Cells product comprises a heterogeneous population of HEK293T cells subjected to CRISPR/Cas9-mediated gene disruption of the ADPGK locus. This polyclonal knockout model provides a physiologically relevant loss-of-function system for studying ADP-dependent glucose phosphorylation and its downstream metabolic consequences without clonal selection bias.
HEK293T cells are a widely used human embryonic kidney epithelial-like cell line stably expressing the SV40 large T antigen. This genetic modification supports episomal replication of plasmids containing the SV40 origin, facilitating high-level transient protein expression and efficient production of lentiviral and retroviral vectors. The cells exhibit robust growth and are amenable to standard culture conditions, making them a versatile platform for genetic perturbation studies in metabolic and oncological research.
ADPGK encodes a glucokinase that uniquely utilizes ADP rather than ATP to phosphorylate glucose to glucose-6-phosphate (G6P), linking glucose metabolism to cellular energy status. Within the glycolytic pathway, ADPGK functions upstream of HK1, PFKL, and LDHA, and its activity is regulated by key stress-responsive transcription factors including HIF1A and TP53. By controlling the entry of glucose into glycolysis and the pentose phosphate pathway, ADPGK influences the production of NADPH and ribose-5-phosphate for antioxidant defense and nucleotide biosynthesis, thereby coupling nutrient availability to biosynthetic and redox homeostasis in proliferating cells.
In HEK293T cells, which rely on rapid glucose consumption to sustain high protein expression and viral packaging demands, ADPGK disruption impairs ADP-dependent glycolytic flux. This metabolic intervention is predicted to attenuate lactate fermentation and compromise NADPH generation via the pentose phosphate pathway, collectively reducing the cell??s capacity to maintain redox balance and support anabolic processes. Consequently, ADPGK knockout cells may exhibit heightened sensitivity to glucose deprivation and metabolic stress, offering a controlled system to interrogate metabolic adaptations and vulnerabilities relevant to tumor cell metabolism.
Researchers can employ this polyclonal knockout population in an array of functional assays to dissect metabolic pathways. Typical applications include western blotting and RT-qPCR to confirm ADPGK loss, metabolic flux analyses measuring glucose uptake, lactate secretion, and G6P levels, as well as quantification of NADPH/NADP+ and ATP/AMP ratios under normoxic or hypoxic conditions. The model is also suited for viability assays under glucose limitation, colony formation studies, and xenograft tumor growth experiments to evaluate the impact of ADPGK on tumorigenicity. For further information or to discuss customized experimental strategies, please contact Ascent Research.
