This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line, featuring targeted disruption of the GPD1L gene. The polyclonal format comprises a heterogeneous pool of cells carrying diverse gene-inactivating edits, creating a robust loss-of-function model without requiring single-cell cloning. This approach enables the study of GPD1L??s roles in cancer metabolism and signaling networks while preserving the inherent genetic diversity of the host cell background.
NCI-H1975 cells are epithelial cells isolated from the pleural effusion of a patient with non-small cell lung cancer (NSCLC) and harbor the clinically relevant EGFR L858R and T790M mutations. The L858R mutation confers constitutive kinase activity, while T790M is a gatekeeper mutation associated with resistance to first-generation EGFR tyrosine kinase inhibitors. This genetic context makes the cell line a widely employed model for investigating EGFR-driven oncogenic signaling, acquired drug resistance, and metabolic adaptations in NSCLC.
GPD1L encodes glycerol-3-phosphate dehydrogenase 1-like protein, a cytosolic enzyme that catalyzes the NADH-dependent reduction of dihydroxyacetone phosphate (DHAP) to glycerol-3-phosphate, linking glycolysis to glycerolipid synthesis. Under the control of HIF1A, PPARG, and glucose, GPD1L-generated glycerol-3-phosphate is directed into triglyceride and phosphatidic acid production. Beyond metabolism, GPD1L directly interacts with the cardiac sodium channel SCN5A and the desmosomal protein PKP2 to modulate channel trafficking and activity; accordingly, GPD1L mutations cause Brugada syndrome and sudden infant death syndrome.
In NCI-H1975 lung cancer cells, GPD1L knockout provides a defined genetic tool to dissect the crosstalk between glycolytic metabolism and lipid synthesis, processes frequently reprogrammed in NSCLC to support rapid proliferation and survival. The presence of EGFR L858R/T790M mutations offers a unique opportunity to examine how GPD1L loss influences metabolic enzyme expression, NADH/NAD+ redox balance, and lipidome remodeling in the context of oncogenic signaling and drug resistance. Moreover, the interaction between GPD1L and SCN5A, though primarily studied in cardiac tissue, may reveal unexpected roles in cancer cell physiology, such as ion channel-mediated regulation of migration or apoptosis.
Researchers can apply this knockout model in a variety of experimental workflows, including Western blotting and RT-qPCR for gene expression analysis, glycerol-3-phosphate and ATP quantification to assess metabolic shifts, and migration or invasion assays to evaluate metastatic potential. Drug sensitivity studies with EGFR inhibitors can elucidate whether GPD1L disruption alters therapeutic responses. High-throughput approaches such as RNA-seq and ChIP-qPCR integrate well with this model to map global transcriptional and epigenetic changes. Additionally, reconstitution experiments with wild-type or mutant GPD1L can validate phenotype specificity. For further information or to inquire about custom editing services, please contact Ascent Research.
