Volohonsky Gloria, Terenzi Olivier, Soichot Julien, Naujoks Daniel A, Nolan Tony, Windbichler Nikolai, Kapps Delphine, Smidler Andrea L, Vittu Anaïs, Costa Giulia, Steinert Stefanie, Levashina Elena A, Blandin Stéphanie A, Marois Eric
INSERM U963, CNRS UPR9022, Université de Strasbourg, Institut de Biologie Moléculaire et Cellulaire, 67084 Strasbourg, France.
Imperial College London, Division of Cell and Molecular Biology, Imperial College Road, London SW7 2AZ, United Kingdom.
G3 (Bethesda). 2015 Apr 13;5(6):1151-63. doi: 10.1534/g3.115.016808.
Transgenesis is an essential tool to investigate gene function and to introduce desired characters in laboratory organisms. Setting-up transgenesis in non-model organisms is challenging due to the diversity of biological life traits and due to knowledge gaps in genomic information. Some procedures will be broadly applicable to many organisms, and others have to be specifically developed for the target species. Transgenesis in disease vector mosquitoes has existed since the 2000s but has remained limited by the delicate biology of these insects. Here, we report a compilation of the transgenesis tools that we have designed for the malaria vector Anopheles gambiae, including new docking strains, convenient transgenesis plasmids, a puromycin resistance selection marker, mosquitoes expressing cre recombinase, and various reporter lines defining the activity of cloned promoters. This toolbox contributed to rendering transgenesis routine in this species and is now enabling the development of increasingly refined genetic manipulations such as targeted mutagenesis. Some of the reagents and procedures reported here are easily transferable to other nonmodel species, including other disease vector or agricultural pest insects.
转基因技术是研究基因功能以及在实验生物中引入所需性状的重要工具。由于生物生命特征的多样性以及基因组信息方面的知识空白，在非模式生物中建立转基因技术具有挑战性。一些方法将广泛适用于许多生物，而其他方法则必须针对目标物种专门开发。自21世纪以来，在病媒蚊子中就存在转基因技术，但一直受到这些昆虫微妙生物学特性的限制。在这里，我们报告了我们为疟疾媒介冈比亚按蚊设计的转基因工具汇编，包括新的对接菌株、方便的转基因质粒、嘌呤霉素抗性选择标记、表达cre重组酶的蚊子，以及定义克隆启动子活性的各种报告系。这个工具箱有助于使该物种的转基因技术成为常规操作，现在还推动了越来越精细的基因操作的发展，如靶向诱变。这里报道的一些试剂和方法很容易转移到其他非模式物种，包括其他病媒或农业害虫昆虫。
