High-throughput enhancer trap by remobilization of transposon Minos in Ciona intestinalis
High-throughput enhancer trap by remobilization of transposon Minos in Ciona intestinalis
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DOI：
10.1002/dvg.20290
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发表时间：
2007-05-01
期刊：
影响因子：
1.5
通讯作者：
Sasakura, Yasunori
中科院分区：
文献类型：
--
作者：
Awazu, Satoko;Matsuoka, Terumi;Sasakura, Yasunori
利用转座子的增强子捕获方法为我们提供了有关基因功能和基因表达模式的信息。在海鞘中，使用Tc 1/mariner转座子Minos实现了基于转座子的转基因和插入突变。我们报道了一种新的技术，增强子陷阱在C。好吧该技术利用了玻璃海鞘基因组中Minos的再动员。构建用于增强子陷阱的Minos载体，并将载体的串联阵列插入引入玻璃海鞘基因组中以产生突变体系。Minos在玻璃海鞘染色体中被重新激活，通过提供转座酶来产生新的插入。这些转座酶导入的动物与野生型动物杂交。近80%的F1家族表现出新的GFP表达模式。这种高通量增强子陷阱筛选将有助于创建新的标记转基因系，显示报告基因在特定组织中的表达，并确定新的基因表达模式。
The enhancer trap approach utilizing transposons yields us information about gene functions and gene expression patterns. In the ascidian Ciona intestinalis, transposon-based transgenesis and insertional mutagenesis were achieved with a Tc1/mariner transposon Minos. We report development of a novel technique for enhancer trap in C. intestinalis. This technique uses remobilization of Minos in the Ciona genome. A Minos vector for enhancer trap was constructed and a tandem array insertion of the vector was introduced into the Ciona genome to create a mutator line. Minos was remobilized in Ciona chromosomes to create new insertions by providing transposases. These transposase-introduced animals were crossed with wild-type animals. Nearly 80% of F1 families showed novel GFP expression patterns. This high-throughput enhancer trap screen will be useful to create new marker transgenic lines showing reporter gene expression in specific tissues and to identify novel patterns of gene expression.