Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster
Post-integration stabilization of a transposon vector by terminal sequence deletion in Drosophila melanogaster
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DOI：
10.1038/nbt1002
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发表时间：
2004-09-01
影响因子：
46.9
通讯作者：
Horn, C
中科院分区：
文献类型：
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作者：
Handler, AM;Zimowska, GJ;Horn, C
近20种昆虫的种系转化系统已经从转座因子中衍生出来，从而允许开发用于基础和应用研究的转基因昆虫（1-3）。这些系统使用有缺陷的非自主载体，在瞬时提供的转座酶辅助质粒消失后，导致稳定的载体整合(4)，这对于维持真正的育种系和一致的转基因表达至关重要，否则在载体重组后就会丢失。到目前为止，实验室研究还没有考虑到意外转座酶来源重新激活的风险，但在生物防治计划中使用数百万转基因昆虫可能会增加风险，因此使其成为野外释放计划的生态安全的关键问题（5,6）。在这里，我们描述了一种有效的方法，删除一个末端重复序列的转座子载体基因组整合后。这个过程防止了转座酶介导的其他末端序列和相关基因的再动员，确保了它们的基因组稳定性。
Germline transformation systems for nearly 20 insect species have been derived from transposable elements, allowing the development of transgenic insects for basic and applied studies(1-3). These systems use a defective nonautonomous vector that results in stable vector integrations after the disappearance of transiently provided transposase helper plasmid(4), which is essential to maintain true breeding lines and consistent transgene expression that would otherwise be lost after vector remobilization. The risk of remobilization by an unintended transposase source has so far not been a concern for laboratory studies, but the prospective use of millions of transgenic insects in biocontrol programs will likely increase the risk, therefore making this a critical issue for the ecological safety of field release programs(5,6). Here we describe an efficient method that deletes a terminal repeat sequence of a transposon vector after genomic integration. This procedure prevents transposase-mediated remobilization of the other terminal sequence and associated genes, ensuring their genomic stability.