Development of serine recombinases as tools for genome engineering in zebrafish

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Site specific recombinases are proteins that promote reciprocal and conservative recombination between specific DNA sequences. In both fruit flies and mice site specific recombinases have been used to turn off genes in particular tissues and to manipulate chromosome structure. This type of manipulation is known as genome engineering. Genome engineering in fruit flies and mice has relied on reversible site specific recombinases. Over the past few years however we have shown that a new type of site specific recombinase; one that promotes a unidrectional or irreversible reaction, is very active in vertebrate cells in cell culture and can be used for elaborate genome engineering reactions in these cells. We would now like to use these enzymes for genome engineering in zebrafish . We wish to establish a methodology for the efficient construction of animals in which particular tissues are labelled with fluorescent proteins and to establish a system for gene targeting by homologous recombination.