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

Bernardini, Federica; Galizi, Roberto; Menichelli, Miriam; Papathanos, Philippos-Aris; Dritsou, Vicky; Marois, Eric; Crisanti, Andrea; Windbichler, Nikolai

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Authors

Federica Bernardini

Miriam Menichelli

Philippos-Aris Papathanos

Vicky Dritsou

Eric Marois

Andrea Crisanti

Nikolai Windbichler



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.

Citation

Bernardini, F., Galizi, R., Menichelli, M., Papathanos, P., Dritsou, V., Marois, E., …Windbichler, N. (2014). Site-specific genetic engineering of the Anopheles gambiae Y chromosome. Proceedings of the National Academy of Sciences of the United States of America, 7600 - 7605. https://doi.org/10.1073/pnas.1404996111

Journal Article Type Article
Acceptance Date Apr 17, 2014
Publication Date May 12, 2014
Publicly Available Date May 26, 2023
Journal PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Print ISSN 0027-8424
Publisher National Academy of Sciences
Pages 7600 - 7605
DOI https://doi.org/10.1073/pnas.1404996111
Keywords Site-specific, genetic, engineering, anopheles gambiae, y chromosome
Publisher URL https://www.pnas.org/content/111/21/7600/tab-article-info
PMID 24821795

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