The ADPGK Knockout A-549 Polyclonal Cells product consists of a CRISPR/Cas9-edited polyclonal knockout cell population derived from the A-549 human lung adenocarcinoma cell line, in which the ADPGK gene has been disrupted to generate a loss-of-function model. This polyclonal population retains genetic heterogeneity and is suitable for studying the collective effects of ADPGK ablation without the bias introduced by single-cell cloning. The use of CRISPR/Cas9-mediated gene disruption ensures efficient targeting of ADPGK, enabling researchers to investigate the metabolic consequences of its inactivation in a relevant epithelial context. The product is provided as a ready-to-use vital frozen stock, designed for immediate propagation and functional analysis.
The host A-549 cell line, established from a human lung adenocarcinoma, is a widely employed model in cancer biology and epithelial cell research. A-549 cells harbor a KRAS mutation (G12S), a common oncogenic driver in non-small cell lung cancer, and grow as an adherent monolayer with an epithelial-like morphology. These cells retain features of alveolar type II pneumocytes, including lamellar body formation and expression of surfactant proteins, making them valuable for studies on lung tumor biology, drug response, and metabolic reprogramming. Their KRAS-mutant background provides a clinically relevant platform to evaluate how ADPGK loss interacts with oncogenic signaling in lung adenocarcinoma.
ADPGK (ADP-dependent glucokinase) encodes a unique enzyme that catalyzes the phosphorylation of glucose to glucose-6-phosphate using ADP as the phosphate donor, bypassing the canonical ATP-dependent hexokinase reaction. This activity contributes to glycolytic flux, especially under conditions of energy stress, and feeds into the pentose phosphate pathway and other metabolic branches. ADPGK is regulated by key factors such as HIF-1??, c-Myc, and mTORC1, and interacts with glycolytic complexes including hexokinase and glucose-6-phosphate isomerase. Downstream, it generates glucose-6-phosphate and other glycolytic intermediates, linking its function to central carbon metabolism. Representative pathway components positioned around ADPGK include hexokinase, phosphoglucose isomerase, phosphofructokinase, aldolase, and pyruvate kinase, highlighting its integration into the glycolytic network.
In the A-549 model, ADPGK knockout eliminates the ADP-dependent glucose phosphorylation route, thereby compromising glycolytic flexibility and reducing metabolic resilience under hypoxia or nutrient deprivation??conditions frequently encountered in the tumor microenvironment. This impairment can suppress tumor cell proliferation and survival, underscoring ADPGK as a potential metabolic vulnerability in KRAS-mutant lung adenocarcinoma. The polyclonal nature of this knockout population allows assessment of heterogeneous responses across a genetically diverse cell pool, closely mimicking the clonal variation seen in patient tumors and providing a more robust platform for target validation than monoclonal isolates.
This product is suited for diverse applications in cancer metabolism research, including investigations into glycolysis inhibition, hypoxia adaptation, and metabolic reprogramming. Typical experimental approaches include measurement of ADPGK activity, lactate production, and glucose uptake; Seahorse metabolic flux analysis; cell viability assays under hypoxic conditions; and molecular readouts via Western blotting, RT-qPCR, or immunofluorescence. The cells enable dissection of ADPGK-dependent mechanisms downstream of HIF-1??, c-Myc, and mTORC1 signaling, and facilitate screening for synthetic lethal interactions or metabolic dependencies. For further details or technical support, please contact Ascent Research.
