Development of serine recombinases as tools for genome engineering in zebrafish

开发丝氨酸重组酶作为斑马鱼基因组工程工具

• 批准号: BB/E000894/1
• 负责人: William Brown
• 金额: $ 45.67万
• 依托单位: University of Nottingham
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2006
• 资助国家: 英国
• 起止时间: 2006 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE000894%2F1
• 关键词: Development; serine; recombinases; tools; genome

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.

位点特异性重组酶是促进特定DNA序列之间的相互和保守重组的蛋白质。在果蝇和小鼠中，位点特异性重组酶已被用于关闭特定组织中的基因并操纵染色体结构。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，在过去的几年中，我们已经表明，一种新型的位点特异性重组酶，一种促进单向或不可逆反应的重组酶，在细胞培养的脊椎动物细胞中非常活跃，并可用于这些细胞中的精细基因组工程反应。我们现在想把这些酶用于斑马鱼的基因组工程。我们希望建立一种有效构建动物的方法，其中特定组织用荧光蛋白标记，并建立一个通过同源重组进行基因靶向的系统。

项目成果

Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of "Rong and Golic".

• DOI: 10.1007/s11248-012-9607-1
• 发表时间: 2012-10
• 期刊: Transgenic research
• 影响因子: 3
• 作者: Brookfield R;Dafhnis-Calas F;Xu Z;Brown W
• 通讯作者: Brown W