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

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