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

一种将精确的基因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至关重要。就蚊子媒介而言，这样的技术可以让我们了解使其成为疟疾寄生虫传播者的特征的遗传本质。世界上每年有一半的人口面临疟疾的风险，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，按蚊是疟疾的重要媒介，在某些地区往往是造成大多数传播的主要物种。尽管如此，在实验室中对它的研究相对较少，部分原因是它很难在实验室中饲养，部分原因是从基因上转化它的技术--即操纵它的基因，看看它们有什么效果--还没有开发出来。该项目将汇集两个研究团队，分别在培育按蚊和蚊子基因转化方面拥有独特和互补的经验。我们将建立新的实验群体，为科学界提供多样化的遗传种群作为资源。我们将使用精确的基因组编辑工具，如CRISPR，它的功能就像一把精确的分子剪刀，可以剪切几乎任何选定的蚊子DNA序列，以在那里引入所需的序列变化。这种技术可以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药性的遗传基础，目前这种抗药性正在危及迄今通过使用驱虫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方式加强蚊子控制：1)量化不同的基因对抗性的贡献；2)分子筛查，以高通量的方式及早发现杀虫剂抗性的出现；3)根据出现的抗性机制，自适应地轮换杀虫剂类别，以延长特定杀虫剂类别的使用时间4)比较按蚊和冈比亚按蚊的抗药性来源。

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