Applications of transgenesis in the chick: developmental studies and disease

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We have shown that lentiviral vectors can be utilised to generate transgenic chickens with efficiencies 10-100-fold higher than established methods. These dramatically improved rates of transgenic animal production are such that many potential applications of transgenesis are now possible that have not been feasible in the past. Our major goal now is to plan and implement projects to exploit these new technologies. The chick is a major model for the study of development in vertebrates and is particularly useful as the embryo in ovo may be easily accessed for a range of manipulations, including electroporation, grafting, application of retroviral vectors and small molecules including morpholinos. Transgenesis may be used to generate new tools for developmental studies and to investigate function of genes involved in development. We have already generated a transgenic line of chickens that express GFP in every cell and demonstrated that embryos of this line may be used in grafting studies with wildtype chick embryos. We have also shown that tissue-specific expression may be obtained from putative regulatory sequences in transgenes delivered by lentiviral vectors. Specific aims include: (1) provision of GFP embryos to the UK chick development community. (2) generation of additional transgenic tools: membrane-localised GFP, photoactivatable GFP for analysis of cell shape in development and single-cell tracking (3) investigation of the function of genes involved in sex determination in birds by gene knockdown and/or overexpression studies.Transgenesis may be used as a tool to study major diseases of poultry and host functions in response to disease. A novel approach we are developing is to produce transgenic animals expressing RNAi transgenes targeted at specific genes of incoming pathogens e.g. expression of a miRNA against two genes of avian influenza and two genes in Marek¿s Disease Virus in chickens.

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