Developing reciprocal chromosomal translocations for wild population replacement in an important vector of human disease.

摘要 这项工作将涉及在寨卡病毒、基孔肯雅病毒和 登革热蚊子Ae.埃及伊蚊是人类昆虫传播疾病的主要媒介，已知每年可感染多达 全世界有5亿人，超过50万人住院，大约25,000人死亡。海流 用于预防蚊子疾病的方法，包括通过环境改造抑制病媒， 杀虫剂和抗炎药根本不够。野生蚊子的更替 具有“抗病”基因改造个体的种群应提供 可持续的、长期的、疾病预防方法。然而，介导疾病的转基因 耐火性不太可能给携带它们的昆虫带来整体健康益处。此外，野生的 种群数量庞大，部分处于生殖隔离状态，分散在大片地区。因此，人口 替换需要一种基因驱动机制来传播连锁的货物基因，从而介导疾病 耐药性，通过野生病原体传播种群。在这里，我提议让历史上的 利用染色体易位将抗病基因传播到野生病原菌的概念 传播蚊子种群。虽然这种方法在过去进行了严格的尝试，但最终 完全被放弃，因为用于生成 易位菌株，除了不能将抗病基因与染色体联系起来外 断点。重要的是，基因工程和合成生物学的最新进展使这些成为可能。 需要彻底克服的历史问题。此外，易位介导的基因驱动系统是 依赖于门槛，因此具有几个吸引人的特征，对于社会和科学接受 野生转基因释放：该系统因物种而异；物种间零水平传播；最低限度 与杀虫剂不同的生态影响；强大而牢不可破，与自私的不可阻挡的联系 基因成分及其货物；如果需要，可以从野生种群中完全移除转基因。 因此，该项目将利用尖端的应用合成生物学原理来设计相互作用 Ae.精确位置的染色体易位埃及(Aim-1)。一旦携带易位的菌株 建立，这些将被引入野生遗传背景，适应度动态将被测量，以及 将进行实验室规模的小型驱动实验(AIM-2)。总体而言，成功的基于移位的 与抗病基因连锁的种群替换系统将对两者产生重大影响 人类健康和未来管理蚊子和其他昆虫的技术能力。 由于这些系统可以在大多数昆虫身上设计，这种创新的方法稍后也可以在 广泛的昆虫病媒，使昆虫种群控制领域发生革命性变化并实现现代化。

Abstract This work will involve the development of an invasive gene drive system in the Zika, Chikungunya, and Dengue mosquito, Ae. aegypti, a major vector of human insect-borne disease known to annually infect up to 500 million people worldwide, hospitalizing over ½ a million, and killing approximately 25,000. The current approaches used for mosquito disease prevention, including vector suppression by environmental modification, insecticides, and anti-inflammatory drugs, are simply insufficient. The replacement of wild mosquito populations with genetically modified individuals that are engineered to be “disease resistant” should provide a sustainable, long-term, method for disease prevention. However, the transgenes that mediate disease refractoriness are unlikely to confer an overall fitness benefit to insects that carry them. Additionally, wild populations are large, partially reproductively isolated, and dispersed over wide areas. Therefore, population replacement requires a gene drive mechanism in order to spread linked cargo genes, mediating disease refractoriness, through wild pathogen transmitting populations. Here I propose to “resurrect” the historical concept of using reciprocal chromosomal translocations to spread disease refractory genes into wild pathogen transmitting mosquito populations. While this approach was rigorously attempted in the past, it was ultimately completely abandoned, due to elevated fitness costs resulting from the technologies used to generate the translocation strains, in addition to the inabilities to link genes for disease resistance to the chromosomal break-points. Importantly, recent advancements in genetic engineering and synthetic biology allow for these historical problems to be entirely overcome. Furthermore, translocation-mediated gene drive systems are threshold-dependent and thus have several attractive features important for social and scientific acceptance for wild transgenic releases: the systems are species specific; zero horizontal spread between species; minimal ecological impact in contrast to insecticides; robust and unbreakable with a inexorable linkage of the selfish genetic element with its cargo; complete transgene removal from wild population can be carried out if desired. Therefore, this project will utilize cutting-edge applied synthetic biology principals to engineer reciprocal chromosomal translocations at precise locations in Ae. aegypti (Aim-1). Once translocation-bearing strains are established, these will be introgressed with wild genetic backgrounds, fitness dynamics will be measured, and small laboratory-scale drive experiments will be executed (Aim-2). Overall, a successful translocation-based population replacement system linked with disease refractory genes will have a significant impact on both human health and the technical capability in which mosquitoes and other insects will be managed in the future. As these systems can be designed in most insects, this innovative approach could also later be engineered in wide range of insect disease vectors, revolutionizing and modernizing the field of insect population control.