Development of serine recombinases as tools for genome engineering in zebrafish

位点特异性重组酶是促进特定DNA序列之间互惠和保守重组的蛋白质。在果蝇和小鼠中，位点特异性重组酶被用于关闭特定组织中的基因和操纵染色体结构。这种类型的操作被称为基因组工程。果蝇和小鼠的基因组工程依赖于可逆位点特异性重组酶。然而，在过去的几年里，我们发现了一种新的位点特异性重组酶；一种促进单向或不可逆反应的基因，在细胞培养的脊椎动物细胞中非常活跃，可以用于这些细胞中精细的基因组工程反应。我们现在想把这些酶用于斑马鱼的基因组工程。我们希望建立一种用荧光蛋白标记特定组织的高效动物构建方法，并建立一种通过同源重组的基因靶向系统。

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.