The ATP7B Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human A-549 lung epithelial cell line. This product enables loss-of-function studies of the ATP7B gene, which encodes a copper-transporting P-type ATPase essential for copper homeostasis. The polyclonal knockout format provides a heterogeneous population of cells with targeted disruption of ATP7B, facilitating functional investigations without the need for single-cell cloning. This gene-edited model is designed for researchers examining copper metabolism, oxidative stress, and related pathophysiological processes in a lung epithelial context.
The A-549 host cell line was originally established from a lung adenocarcinoma of a 58-year-old Caucasian male and serves as a widely used model for alveolar type II epithelial cells. These cells retain key features of lung adenocarcinoma, including epithelial morphology, growth characteristics, and oncogenic signaling pathways. Their epithelial origin makes them suitable for studying copper transport mechanisms in pulmonary epithelial biology and cancer. The A-549 background provides a well-characterized platform for investigating the role of ATP7B in copper homeostasis within a tumorigenic environment.
ATP7B functions as a copper-transporting ATPase that primarily mediates biliary copper excretion and the incorporation of copper into ceruloplasmin. Under normal conditions, ATP7B traffics from the trans-Golgi network to the plasma membrane in response to elevated intracellular copper, facilitating copper efflux. Its activity is regulated by copper-bound ATOX1 chaperone, COMMD1, and the transcription factor MTF1, while downstream targets include ceruloplasmin, superoxide dismutase 3 (SOD3), and lysyl oxidase (LOX). The mechanistic summary provided with this product indicates that knockout of ATP7B impairs copper export and ceruloplasmin loading, leading to intracellular copper accumulation, increased reactive oxygen species (ROS) production, and diminished cuproenzyme activity.
In the A-549 lung adenocarcinoma context, ATP7B disruption induces copper accumulation that can exacerbate oxidative stress and modulate tumor cell behavior. The lung epithelial environment is particularly sensitive to redox imbalance, and ATP7B loss may alter responses to environmental copper exposure and chemotherapeutic agents. By linking copper homeostasis to oxidative stress pathways involving HIF1A, this knockout model enables dissection of how copper dysregulation influences lung cancer cell survival, proliferation, and metastasis. It also provides a tractable system for exploring the interplay between copper metabolism and oncogenic signaling in epithelial cells.
This polyclonal knockout cell population is suitable for a wide range of research applications, including Wilson disease modeling, studies on copper homeostasis and metal ion transport, oxidative stress investigations, and drug screening for copper chelators. Moreover, it serves as a valuable tool for cancer copper biology and lung adenocarcinoma research. Representative assays that can be performed include Western blotting, RT-qPCR, copper quantification, ceruloplasmin ferroxidase activity measurement, ROS detection, cell viability under copper challenge, immunofluorescence staining, and Sanger sequencing for knockout verification. For technical inquiries or additional product information, please contact Ascent Research.
