The ATP7B Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for the targeted disruption of the ATP7B gene in the Jurkat T-lymphocyte background. This polyclonal loss-of-function model is produced by introducing CRISPR/Cas9 components to generate a heterogeneous pool of edited cells, abolishing wild-type ATP7B expression without selection for a single clonal genotype. The product provides a robust cellular system for investigating copper homeostasis pathways and associated disease mechanisms in an immune cell context. Researchers can utilize these cells to probe ATP7B-dependent processes, including copper trafficking, oxidative stress responses, and cuproenzyme maturation, in a well-established T-cell line.
Jurkat cells are an immortalized human T-lymphocyte line derived from the peripheral blood of a 14-year-old male with acute T-cell leukemia. The E6-1 clone, widely adopted in immunology and cancer biology, serves as a premier model for studying T-cell receptor signaling, apoptosis, and cytokine production due to its rapid growth and genetic tractability. This host cell line retains key aspects of T-cell physiology, making it particularly suitable for examining how copper dysregulation affects immune cell function. The Jurkat background thus enables investigation of ATP7B’s role in a lineage outside the classic hepatic context, offering insights into copper metabolism in lymphoid cells.
ATP7B encodes a copper-transporting P-type ATPase that maintains intracellular copper homeostasis by exporting copper from the cytosol into the trans-Golgi network for incorporation into cuproenzymes and across the plasma membrane for excretion. Mechanistically, ATP7B cycles between the Golgi and plasma membrane in a copper-dependent manner, regulated upstream by copper ions, the transcription factor TFEB, and retinoic acid. It interacts directly with the copper chaperone ATOX1 to receive copper and delivers it to target proteins such as ceruloplasmin, a key ferroxidase. Downstream, ATP7B activity supports the function of superoxide dismutase 1 (SOD1) and overall cellular copper efflux, while its activity is modulated by the COMMD1 protein. Disruption of ATP7B leads to copper accumulation, elevated oxidative stress, and impaired cuproenzyme activity, mirroring the molecular defects observed in Wilson disease.
In the Jurkat T-cell context, ATP7B knockout creates a unique model to study copper-induced oxidative stress and apoptosis in immune cells. T lymphocytes depend on exquisite copper regulation for proliferative signaling and redox balance, and loss of ATP7B function disrupts this equilibrium, sensitizing cells to copper toxicity. This model allows dissection of how copper overload modulates T-cell receptor pathways, mitochondrial function, and apoptotic cascades, providing a surrogate system for Wilson disease research beyond hepatocytes. The polyclonal nature ensures a diverse range of genotypes, more closely mimicking the heterogeneous genetic lesions encountered in disease states and in drug screening populations.
These polyclonal knockout cells are ideally suited for applications such as exploring copper homeostasis in T cells, modeling molecular defects of Wilson disease in non-hepatic cells, studying copper-induced oxidative stress and apoptosis, and screening for modulators of copper toxicity. Representative assays include western blotting for ATP7B and cuproenzymes, flow cytometric analysis of apoptosis and reactive oxygen species, inductively coupled plasma mass spectrometry for total copper quantification, ceruloplasmin ferroxidase activity measurements, immunofluorescence to assess ATP7B localization, and cell viability assays under copper challenge. For additional information or custom requirements, please contact Ascent Research.
