The role of the circadian clock in the behavior of the malaria mosquito Anopheles coluzzii

项目摘要 科氏按蚊是撒哈拉以南非洲最重要的媒介之一。其中一个原因是它的高 对人类宿主的偏好像所有的生物一样，一个。coluzzii有一个内部时钟， 生物学和行为例如，An. coluzzii是活跃在傍晚和夜间，交配，主机 在这段时间里，寻找、吃糖和产卵都发生了。例如，自发的飞行活动 这种蚊子在白天几乎不存在，但在夜间显示出两个强烈的活动高峰： 一个是傍晚，另一个是深夜。以前的工作也表明，表达式 许多基因都是由生物钟控制的。这包括与嗅觉、免疫和 杀虫剂抗药性内部时钟由几个基因组成，包括基因时钟和周期， 是有节奏地表达的。与其他基因一起，它们形成一个反馈回路， 每24小时一次在夜间活动的伊蚊中， 周期基因导致宿主寻找减少、发育时间延长和存活率降低。 由于体内生物钟对白天和夜间蚊子的影响不同， 不能外推到按蚊身上因此，为了检查在多大程度上关闭内部时钟， 影响着安的行为、发育和生存。coluzzii，我们将使用CRISPR-Cas9来禁用时钟 或周期基因。我们将评估这如何影响自发飞行活动，宿主寻找，交配成功， 以及发育率和存活率。这将告诉我们这种蚊子的行为在多大程度上受到控制 通过内部时钟与外部条件（例如亮/暗）。它也可能提供一种方法来改变蚊子 行为，例如通过减少宿主寻找，以降低其传播疟疾的能力。

Project Summary Anopheles coluzzii is one of most important vectors in sub-Saharan Africa. One reason for this is its high preference for human hosts. Like all organisms, An. coluzzii has an internal clock that controls much of its biology and behavior. For example, An. coluzzii is active in the evening and at night, and mating, host seeking, sugar feeding and oviposition takes places during this time. For example, spontaneous flight activity of this mosquito is all but absent during the day but shows two strong peaks of activity during the night: one in the early evening and one towards the end of the night. Previous work has also shown that the expression of many genes is controlled by the internal clock. This includes genes involved in olfaction, immunity and insecticide resistance. The internal clock consists of several genes, including the genes clock and cycle, that are expressed rhythmically. Together with other genes they form a feedback loop that cycles approximately every 24 hours. In Aedes mosquitoes, which are active at night, shutting down this internal clock by disabling the cycle gene results in much reduced host seeking, longer development times and reduced survival. Because the internal clock affects day-time and night-time mosquitoes differently, the results from Aedes cannot be extrapolated to Anopheles. Therefore, to examine to what extent shutting down the internal clock affects the behavior, development and survival of An. coluzzii, we will use CRISPR-Cas9 to disable the clock or cycle gene. We will evaluate how this affects spontaneous flight activity, host seeking, mating success, as well as developmental rate and survival. This will tell us to what extent this mosquito’s behavior is controlled by the internal clock vs external conditions (e.g light/dark). It may also provide a way to modify mosquito behavior, e.g. by reducing host seeking, to reduce its ability to transmit malaria.