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Silencing the buzz: a new approach to population suppression of mosquitoes by feeding larvae double-stranded RNAs

机译:降低嗡嗡声:通过喂养幼虫双链RNA抑制蚊子种群的新方法

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Background Mosquito-borne diseases threaten over half the world’s human population, making the need for environmentally-safe mosquito population control tools critical. The sterile insect technique (SIT) is a biological control method that can reduce pest insect populations by releasing a large number of sterile males to compete with wild males for female mates to reduce the number of progeny produced. Typically, males are sterilized using radiation, but such methods can reduce their mating competitiveness. The method is also most effective if only males are produced, but this requires the development of effective sex-sorting methods. Recent efforts to use transgenic methods to produce sterile male mosquitoes have increased interest in using SIT to control some of our most serious disease vectors, but the release of genetically modified mosquitoes will undoubtedly encounter considerable delays as regulatory agencies deal with safety issues and public concerns. Methods Testis genes in the dengue vector Aedes aegypti were identified using a suppression subtractive hybridization technique. Mosquito larvae were fed double-stranded RNAs (dsRNAs) that targeted both the testis genes and a female sex determination gene (doublesex) to induce RNA interference (RNAi) -mediated sterility and inhibition of female development. Fertility and mating competiveness of the treated males were assessed in small-scale mating competition experiments. Results Feeding mosquito larvae dsRNAs targeting testis genes produced adult males with greatly reduced fertility; several dsRNAs produced males that were highly effective in competing for mates. RNAi-mediated knockdown of the female-specific isoform of doublesex was also effective in producing a highly male-biased population of mosquitoes, thereby overcoming the need to sex-sort insects before release. Conclusions The sequence-specific gene-silencing mechanism of this RNAi technology renders it adaptable for species-specific application across numerous insect species. We envisage its use for traditional large-scale reared releases of mosquitoes and other pest insects, although the technology might also have potential for field-based control of mosquitoes where eggs deposited into a spiked larval site lead to the release of new sterile males.
机译:背景技术蚊媒疾病威胁着全球一半以上的人口,因此迫切需要对环境安全的蚊子种群控制工具的需求。不育昆虫技术(SIT)是一种生物防治方法,可以通过释放大量不育雄性雄性与野生雄性竞争雌性同性异体来减少所产生的后代数量,从而减少害虫的数量。通常,使用辐射对雄性动物进行消毒,但是这种方法会降低其交配竞争力。如果只生产雄性,该方法也是最有效的,但这需要开发有效的性别分类方法。最近使用转基因方法生产不育雄性蚊子的努力使人们越来越有兴趣使用SIT控制我们一些最严重的病媒,但是转基因蚊子的释放无疑会遇到相当大的延误,因为监管机构要处理安全问题和公众关注。方法采用抑制消减杂交技术鉴定登革热伊蚊的睾丸基因。蚊子幼虫被喂食以睾丸基因和雌性决定基因(doublesex)为目标的双链RNA(dsRNA),以诱导RNA干扰(RNAi)介导的不育和抑制女性发育。在小规模的交配竞争实验中评估了处理过的雄性的生育力和交配能力。结果喂食针对睾丸基因的蚊虫幼虫dsRNA,可以使成年男性的生育能力大大降低。几种dsRNA产生了在竞争配偶方面非常有效的雄性。 RNAi介导的雌雄同体的双性恋基因敲除在产生高度男性偏向的蚊子种群方面也很有效,从而克服了在释放昆虫之前对昆虫进行性别分类的需求。结论该RNAi技术的序列特异性基因沉默机制使其适用于多种昆虫物种的物种特异性应用。我们设想将其用于传统的大规模诱捕蚊子和其他害虫的释放,尽管该技术也可能具有现场控制蚊子的潜力,在这种情况下,沉积在尖峰幼虫部位的卵会导致新的不育雄性动物的释放。

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