Opening up Anopheles funestus to functional genetics and the study of insecticide resistance

一种将精确的遗传变化引入生物体的方法对于理解基因如何决定关键的生物学特征是必不可少的。就蚊子载体而言，这种技术可以让我们了解使其成为疟疾寄生虫如此有弹性的传播者的性状的遗传性质。世界上每年有一半人口面临疟疾的威胁，尽管最近的干预措施取得了一些成功，但仍有近50万人死亡，其中大多数是儿童。在撒哈拉以南非洲的许多地区，狐按蚊是疟疾的一个重要媒介，在某些地区往往是造成大多数传播的主要物种。尽管如此，它在实验室中的研究相对较少，部分原因是它很难在实验室中饲养，部分原因是一种基因改造技术——即操纵它的基因来观察它们有什么影响——尚未开发出来。该项目将汇集两个研究小组，他们分别在养疟蚊和蚊子遗传转化方面具有独特和互补的经验。我们将建立新的实验室菌落，为科学界提供不同的遗传群体作为资源。我们将使用精确的基因组编辑工具，比如CRISPR，它可以像一把精确的分子剪刀一样，剪切几乎任何蚊子的DNA序列，从而在那里引入所需的序列变化。这种技术允许确认基因序列的作用，这些基因序列以前可能与决定这种蚊子传播疟疾能力的重要特征有关。这项研究建议的重点将是了解杀虫剂抗性的遗传基础，这种抗性目前正在危及迄今为止通过使用经杀虫剂处理的蚊帐和室内喷洒杀虫剂所取得的成果。对抗性机制的了解将通过以下途径加强蚊虫控制：1)对抗性的不同遗传贡献进行量化；2)分子筛选，早期发现抗药性，高通量检测；3)针对抗性机制的适应性杀虫剂类别轮作，以延长特定杀虫剂类别的效用。4)比较富氏按蚊和冈比亚按蚊杀虫剂抗性的来源

A way to introduce precise genetic changes into an organism is essential for understanding how genes determine key biological characteristics. In the case of mosquito vectors such a technology can allow us to understand the genetic nature of traits that make it such a resilient transmitter of the malaria parasite. Half the world's population are at risk from malaria every year and, despite recent interventions that have had some success, close to half a million people die, most of these children. Anopheles funestus is a significant vector of malaria in many areas of sub-Saharan Africa, often being the dominant species responsible for the majority of transmission in some areas. Despite this, it is relatively poorly studied in the lab, in part because it can be difficult to rear in the laboratory, in part because a technology to genetically transform it - i.e. manipulate its genes to see what effect they have - has yet to be developed. This project will bring together two research teams with unique and complementary experience in the rearing of Anopheles funestus and in genetic transformation of mosquitoes, respectively. We will establish new laboratory colonies of A. funestus, providing varied genetic populations as resource for the scientific community. We will use precise genome editing tools, such as CRISPR, which can function like a precise pair of molecular scissors to cut almost any mosquito DNA sequence of choice to introduce desired sequence changes there. Such a technology permits confirmation of the role of genetic sequences that might previously have been associated with important traits that determine the ability of this mosquito to transmit malaria. The focus of this research proposal will be to understand the genetic basis of insecticide resistance, which is currently jeopardising the gains achieved to date by using insecticide-treated nets and indoor spraying of insecticides. An understanding of the mechanism of resistance will enhance mosquito control through: 1) allowing a quantification of the different genetic contributions to resistance; 2) molecular screening to detect early the emergence of insecticide resistance and in a high throughput fashion; 3) adaptive rotation of insecticide classes in response to mechanism of resistance emerging, in order to prolong the utility of a given insecticide class 4) a comparison of the origins of insecticide resistance among Anopheles funestus and Anopheles gambiae