Abstract
BackgroundGermline specific promoters are an essential component of potential vector control strategies which function by genetic drive, however suitable promoters are not currently available for the main human malaria vector Anopheles gambiae.ResultsWe have identified the Anopheles gambiae vasa-like gene and found its expression to be specifically localized to both the male and female gonads in adult mosquitoes. We have functionally characterised using transgenic reporter lines the regulatory regions required for driving transgene expression in a pattern mirroring that of the endogenous vasa locus. Two reporter constructs indicate the existence of distinct vasa regulatory elements within the 5' untranslated regions responsible not only for the spatial and temporal but also for the sex specific germline expression. vasa driven eGFP expression in the ovary of heterozygous mosquitoes resulted in the progressive accumulation of maternal protein and transcript in developing oocytes that were then detectable in all embryos and neonatal larvae.ConclusionWe have characterized the vasa regulatory regions that are not only suited to drive transgenes in the early germline of both sexes but could also be utilized to manipulate the zygotic genome of developing embryos via maternal deposition of active molecules. We have used computational models to show that a homing endonuclease-based gene drive system can function in the presence of maternal deposition and describe a novel non-invasive control strategy based on early vasa driven homing endonuclease expression.
Highlights
Germline specific promoters are an essential component of potential vector control strategies which function by genetic drive, suitable promoters are not currently available for the main human malaria vector Anopheles gambiae
We show that the activity of this Anopheles regulatory region depends on distinct elements and that such activity is suitable for the expression of transgenes in both the male and the female germline prior to meiosis, and allows us to propose a novel inundative genetic control strategy that combines the advantages of genetic drive and dominant sterility
These often rely on the ability of the control trait to increase in frequency once released, a feature that is inherently determined by its function in the mosquito germlines
Summary
Germline specific promoters are an essential component of potential vector control strategies which function by genetic drive, suitable promoters are not currently available for the main human malaria vector Anopheles gambiae. The translation of any such achievements into suitable control measures would require the development of strategies to spread the genetic modification from few laboratory reared insects to large wild type populations, since population replacement strategies based on the massive release of genetically modified mosquitoes carrying a desired trait such as malaria refractoriness are obviously difficult to implement This represents a major scientific and technical challenge. We show that the activity of this Anopheles regulatory region depends on distinct elements and that such activity is suitable for the expression of transgenes in both the male and the female germline prior to meiosis, and allows us to propose a novel inundative genetic control strategy that combines the advantages of genetic drive and dominant sterility
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