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Proc Natl Acad Sci U S A. 2014 May 27;111(21):7600-5. doi: 10.1073/pnas.1404996111. Epub 2014 May 12.

Site-specific genetic engineering of the Anopheles gambiae Y chromosome.

Author information

1
Department of Life Sciences, South Kensington Campus, Imperial College London, London SW7 2AZ, United Kingdom;
2
Dipartimento di Medicina Sperimentale Via Gambuli, Centro di Genomica Funzionale, University of Perugia, 06132 Perugia, Italy; and.
3
Institut de Biologie Moléculaire et Cellulaire, Institut National de la Santé et de la Recherche Médicale U963, Centre National de la Recherche Scientifique, Unité Propre de Recherche 9022, 67084 Strasbourg, France.
4
Department of Life Sciences, South Kensington Campus, Imperial College London, London SW7 2AZ, United Kingdom;Dipartimento di Medicina Sperimentale Via Gambuli, Centro di Genomica Funzionale, University of Perugia, 06132 Perugia, Italy; and.
5
Department of Life Sciences, South Kensington Campus, Imperial College London, London SW7 2AZ, United Kingdom; n.windbichler@imperial.ac.uk.

Abstract

Despite its function in sex determination and its role in driving genome evolution, the Y chromosome remains poorly understood in most species. Y chromosomes are gene-poor, repeat-rich and largely heterochromatic and therefore represent a difficult target for genetic engineering. The Y chromosome of the human malaria vector Anopheles gambiae appears to be involved in sex determination although very little is known about both its structure and function. Here, we characterize a transgenic strain of this mosquito species, obtained by transposon-mediated integration of a transgene construct onto the Y chromosome. Using meganuclease-induced homologous repair we introduce a site-specific recombination signal onto the Y chromosome and show that the resulting docking line can be used for secondary integration. To demonstrate its utility, we study the activity of a germ-line-specific promoter when located on the Y chromosome. We also show that Y-linked fluorescent transgenes allow automated sex separation of this important vector species, providing the means to generate large single-sex populations. Our findings will aid studies of sex chromosome function and enable the development of male-exclusive genetic traits for vector control.

KEYWORDS:

SIT; biotechnology

PMID:
24821795
PMCID:
PMC4040617
DOI:
10.1073/pnas.1404996111
[Indexed for MEDLINE]
Free PMC Article

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