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The aim of the project is to use embryonic stem (ES) cell technology to create chickens that produce novel therapeutic protein in eggs. Somatic and germline chimeras can be generated by injecting freshly isolated cells from stage X embryos into recipient embryos of the same stage.
Efficiency is reduced when chicken ES cells are injected. Our specific aim is to increase the frequency of high-grade and germline chimeras by maximizing the incorporation of ES cells. In phase I, chicken ES cells expressing green fluorescent protein (GFP) will be used as a reporter system to evaluate (a) the suitability of stage VIII and IX recipients embryos from premature eggs and (b) the relationship between the duration in culture of the cES cell line and it's ability to contribute to the germline. The embryos will be incubated using Origen's established surrogate shell system, modifying the conditions to suit the premature eggs if necessary. The options that give the maximum number of viable somatic chimeras and the highest level of germline transmission will be adopted in phase II to establish flocks of chickens that deposit novel proteins in their eggs.
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A hen manufactures several grams of protein daily in egg white and yolk. Origen will harness this protein production system by creating genetically engineered chickens using ES cell technology. Chimeras will be generated using cES cells carrying human genes coding for a range of therapeutic proteins, including monoclonal and polyclonal antibodies. Within 20 weeks of hatching, these novel proteins will be deposited in eggs from high-grade chimeras, and germ-line chimeras will founder transgenic flocks enabling large-scale, cost-effective production of high-quality therapeutic proteins.