Production of High-Grade Germline Chimeras for Genetic Manipulation of Chickens

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.