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High efficiency germ-line transformation of mosquitoes

Nature Protocols volume 1, pages 1312–1317 (2006)Cite this article

Abstract

The ability to manipulate the mosquito genome through germ-line transformation provides us with a powerful tool for investigating gene structure and function. It is also a valuable method for the development of novel approaches to combating the spread of mosquito-vectored diseases. To date, germ-line transformation has been demonstrated in several mosquito species. Transgenes are introduced into pre-blastocyst mosquito embryos using microinjection techniques that take a few hours, and progeny are screened for the presence of a marker gene. The microinjection protocol presented here can be applied to most mosquitoes and contains several improvements over other published methods that increase the survival of injected embryos and, therefore, the number of transformants. Transgenic lines can be established in approximately 1 month using this technique.

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Figure 1
Figure 2: Development and melanization of Anopheles gambiae and Aedes aegypti embryos.
Figure 3: Aedes aegypti embryos lined up for injection.
Figure 4: Schematic of embryo being injected.

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Acknowledgements

The authors would like to thank Sandi Kennedy for help in preparing the manuscript.

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Authors and Affiliations

  1. Department of Biological Sciences, Center for Global Health and Infectious Diseases, University of Notre Dame, Notre Dame, 46556, IN, USA

    Neil F Lobo, Malcolm J Fraser & Frank H Collins

  2. Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, 21205, MD, USA

    John R Clayton

  3. Division of Cell & Molecular Biology, Imperial College of Science, Technology and Medicine, South Kensington Campus, Sir Alexander Fleming Building, London, SW7 2AZ, UK

    Fotis C Kafatos

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  1. Neil F Lobo
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Correspondence to Frank H Collins.

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Lobo, N., Clayton, J., Fraser, M. et al. High efficiency germ-line transformation of mosquitoes. Nat Protoc 1, 1312–1317 (2006). https://doi.org/10.1038/nprot.2006.221

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