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An Anopheles transgenic sexing strain for vector control

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Abstract

Genetic manipulation of mosquito species that serve as vectors for human malaria is a prerequisite to the implementation of gene transfer technologies for the control of vector-borne diseases. Here we report on the development of transgenic sexing lines for the mosquito Anopheles stephensi, the principal vector of human malaria in Asia. Male mosquitoes, expressing enhanced green fluorescent protein (EGFP) under the control of the β2-tubulin promoter, are identified by their fluorescent gonads in as early as their 3rd instar larval stage, and can be efficiently separated from females using both manual methods and automated sorting machines. Importantly, β2-EGFP males are not impaired in their mating ability and viable fluorescent spermatozoa are also detected in spermathecae of wild-type females mated with transgenic males. The transgenic mosquito lines described here combine most of the features desired and required for a safe application of transgenic methodologies to malaria-control programs.

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Acknowledgements

We wish to thank Charles Godfray and Chris Curtis for helpful suggestions, Elisa Petris and Ann Cronin for help with the injections, and Oliver Billker for help with the microphotographs. We also wish to thank Rico Boongarts from Union Biometrica for assisting us in the automated sorting experiments. The project was supported by the Wellcome Trust and BBSRC. F.C. was sponsored by the Wellcome Trust, J.P.B. by the FP6-EU BioMolPar program.

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Competing interests

The authors declare no competing financial interests.

Correspondence to Andrea Crisanti.

Supplementary information

  1. Supplementary Fig. 1

    Schematic representation of transformation vector pPB[DsRed]β2EGFP. (PDF 91 kb)

  2. Supplementary Table 1

    β2-EGFP expression as a marker to predict sex. (PDF 64 kb)

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Figure 1: Fluorescent microphotographs showing β2-EGFP expression at different developmental stages.
Figure 2: Automated sorting of transgenic male larvae.
Figure 3: Dissected spermathecae of wild-type A. stephensi females mated to transgenic heterozygous β2-EGFP males from line β2-egfp2, containing multiple insertions.
Figure 4: Mate choice experiment between transgenic β2-egfp2 and wild-type A. stephensi males.