Abstract
Conditional cell death systems are useful for various aspects of basic science with a wide range of applications, including genetic pest control. We recently demonstrated that expression of the mammalian pro-apoptotic factor, B-cell leukaemia/lymphoma 2-associated X protein (Bax), can induce apoptosis in specific tissues by using tissue specific promoters in silkworm and mosquito. Here, we newly identified a functional promoter in the Asian malaria vector, Anopheles stephensi, which enables gene expression specifically in the testis. We produced a transgenic mosquito line that expresses mouse Bax under the control of this testis-specific promoter. Transgenic mosquito males exhibited aberrant testes without functional sperm and complete sterility, whereas transgenic females maintained normal fecundity. Despite their abnormal testes, the transgenic males maintained normal function of male accessory glands and typical mating behaviour. As a result of mating with these males, females showed refractoriness to further mating. These results suggest that transgenic males induce female sterility via mating. The mosquito is one of the most important disease vectors, and the control of their population benefits global public health. Thus, this Bax-mediated synthetic male-specific sterilization system could be applied to population control of mosquitoes.
Highlights
Insect pests are detrimental to health, agricultural production, and economic progress worldwide
B-cell leukaemia/lymphoma 2-associated X protein (Bax) and Bak (Bcl-2 homologues antagonistic killer) form oligomers resulting in pores on the mitochondrial outer membrane, a process known to be involved in apoptosis and the potentiation of interferon response through mtDNA efflux[25,26,34]
We newly identified a testis-specific promoter in An. stephensi, and established lines expressing mouse B-cell leukaemia/lymphoma 2-associated X protein gene (mBax) under the control of this promoter
Summary
Insect pests are detrimental to health, agricultural production, and economic progress worldwide. Homing CRISPR/Cas9-based gene drives still need improvement[12,13] Another genetic population control strategy is the release of sterilized mosquito males to reduce the size of field populations, which known as the sterile insect technique (SIT). It was reported that expression system using Cas[9] under the control of b2t promoter and a guide RNA that targeted the X-linked rDNA repeat sequences during spermatogenesis was applicable for induction of X-chromosome shredding and production of the male-biased progeny[23]. This X-chromosome shredding-based gene-drive system will be effective reducing the population. MBax could be a useful tool for mosquito control as well as overall insect management
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