Novel transgenic technology to study development in the chick embryo

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We have developed a method for production of transgenic chickens using lentiviral vectors (McGrew et al., 2004) and have used this method to develop lines of transgenic hens synthesising therapeutic proteins as a component of egg white (Lillico et al., 2007). We now aim to develop transgenic techniques to increase the utility of the chick embryo for the study of vertebrate development. The recently described method for culture of primordial germ cells (PGCs) from chick embryos (van de Lavoir et al., 2006) has the potential to form the basis of the development of methods for genetic modification in the chick and also for studies on the cell and developmental biology of germ cell development in vertebrates. A key attribute of the cultured PGCs is that, if the culture conditions are altered, they can change status and become ES cell-like. These cells can then be used to make chimeric embryos. This feature allows analysis of genetically-modified PGCs in the developing embryo, without the time-consuming and expensive requirement to establish transgenic birds. The aim of this project is to utilise our inhouse expertise in the use of lentiviral vectors for transgenesis, transgene design and embryo manipulation to utilise the cultured PGCs for transgenesis and exemplify the use of genetically-modified PGCs in developmental studies. As this method is very new there are many ways in which it can be developed. This PhD project will involve (1) development of basic methods for transgenic manipulation (2) using GM PGCs to study transgene expression in ES cells derived from PGCs and chimeric embryos derived from the ES cells.

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