piggyBac transposon remobilization and enhancer detection in Anopheles mosquitoes
piggyBac transposon remobilization and enhancer detection in Anopheles mosquitoes
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DOI：
10.1073/pnas.1110628108
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发表时间：
2011-09-27
影响因子：
11.1
通讯作者：
Harrell, Robert A., II
中科院分区：
文献类型：
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作者：
O'Brochta, David A.;Alford, Robert T.;Harrell, Robert A., II
蚊子生物学的技术进步使病媒控制的新方法得以发展。缺乏强大的前向遗传学技术，如增强子和基因陷阱，这些技术允许从转座子插入突变引起的表型中确定基因功能。我们发现piggyBac转座子在人类疟疾媒介斯氏按蚊的种系中高度活跃。当从第二个整合的转基因中提供piggyBac转座酶时，含有单个6kb piggyBac元件和表达EGFP标记基因的转基因史蒂芬氏杆菌的后代中，高达6%的人在新的基因组位置上具有载体。piggyBac的主动转位导致了增强子的有效检测，在3龄和4龄幼虫和成虫中，大约10%的piggyBac后代在新的位置具有新的EGFP表达模式。先进的转基因技术如高效的基于转座子的按蚊前向遗传技术的应用，不仅将加速我们对蚊子功能基因组学的理解，以及新的媒介和疾病传播控制策略的发展，而且还将促进进化发育生物学家、病毒学家和寄生虫学家的研究。
Technical advances in mosquito biology are enabling the development of new approaches to vector control. Absent are powerful forward-genetics technologies, such as enhancer and gene traps, that permit determination of gene functions from the phenotypes arising from transposon insertion mutations. We show that the piggyBac transposon is highly active in the germline of the human malaria vector Anopheles stephensi. Up to 6% of the progeny from transgenic A. stephensi containing a single 6-kb piggyBac element with a marker gene expressing EGFP had the vector in new genomic locations when piggyBac transposase was provided in trans from a second integrated transgene. The active transposition of piggyBac resulted in the efficient detection of enhancers, with similar to 10% of the progeny with piggyBac in new locations with novel patterns of EGFP expression in third and fourth instar larvae and in adults. The availability of advanced transgenic capabilities such as efficient transposon-based forward-genetics technologies for Anopheles mosquitoes not only will accelerate our understanding of mosquito functional genomics and the development of novel vector and disease transmission control strategies, but also will enable studies by evolutionary developmental biologists, virologists, and parasitologists.