The GPD1L Knockout A-549 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal cell population derived from the human A-549 lung adenocarcinoma epithelial cell line, in which the GPD1L gene has been disrupted using CRISPR/Cas9. This heterogeneous knockout pool preserves the diversity of editing outcomes, providing a robust loss-of-function model that avoids clonal artifacts and enables population-level functional studies. The product is supplied as a viable polyclonal knockout population, ready for expansion and downstream experiments.
A-549 cells are an established model of human lung adenocarcinoma, originally isolated from a 58-year-old male. They display characteristic epithelial morphology and express wild-type p53, making them a valuable platform for investigating cancer cell biology, signal transduction, and metabolic regulation. Their adherent growth and well-characterized baseline properties facilitate consistent experimental manipulation, including transfection, drug treatment, and metabolic perturbation, thereby offering a physiologically relevant context for interrogating GPD1L function in cancer.
GPD1L encodes a glycerol-3-phosphate dehydrogenase that participates in the glycerol-3-phosphate shuttle, linking cytosolic glycolysis to mitochondrial oxidative phosphorylation. It modulates the intracellular NADH/NAD+ ratio, regulating the activity of the cardiac sodium channel SCN5A. Upstream regulators include the NADH/NAD+ redox couple, cAMP-PKA signaling, PPAR??, and HIF1??. Downstream, GPD1L influences SCN5A gating, GPD1-mediated glycerophospholipid metabolism, and the regeneration of NAD+ from dihydroxyacetone phosphate. Interacting partners such as NDUFS1 (complex I) and NME1 further integrate GPD1L into mitochondrial electron transport and nucleotide metabolism. Representative pathway components span SCN5A, GPD1, the NADH shuttle, and signaling kinases PKC and PKA.
In the A-549 background, GPD1L disruption perturbs redox homeostasis and may alter ion channel function and metabolic reprogramming relevant to both cardiac pathophysiology and cancer. The presence of wild-type p53 permits investigation of p53-dependent metabolic interactions with GPD1L, while the lung adenocarcinoma origin allows dissection of tumor-specific roles. This model is therefore suited for studying the intersection of NAD metabolism, ion channel regulation, and cancer cell behavior, offering insights into Brugada syndrome mechanisms in a non-cardiac context and the potential involvement of GPD1L in tumorigenesis.
Typical applications include metabolic flux analysis using Seahorse assays, NAD/NADH ratio measurements, and MTT viability assays to assess redox-dependent growth effects. Patch-clamp electrophysiology enables direct evaluation of SCN5A activity modulation, supporting ion channel modulator screening. Western blotting, RT-qPCR, and immunofluorescence facilitate validation of knockout and downstream target expression. Migration assays can probe GPD1L??s role in cancer cell motility. Researchers may also employ these cells to explore HIF1??- or PPAR??-mediated regulation. For further details, please contact Ascent Research.
