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 登革蚊Ae.埃及伊蚊是人类虫媒疾病的主要媒介，已知每年感染 全世界有5亿人，超过50万人住院，大约25000人死亡。当前 用于预防蚊子疾病的方法，包括通过改变环境抑制病媒， 杀虫剂和消炎药根本不够。野生蚊子的替代 具有被改造成“抗病”的转基因个体的群体应该提供 可持续、长期的疾病预防方法。然而，介导疾病的转基因 不应性不太可能赋予携带它们的昆虫整体适合性益处。此外，野生 种群庞大，部分生殖隔离，并分散在广阔的地区。因此，人口 替换需要基因驱动机制，以传播连接的货物基因，介导疾病 通过野生病原体传播种群。在这里，我建议“复活”历史 利用相互染色体易位将疾病抗性基因传播到野生病原体中的概念 传播蚊子种群。虽然这种方法在过去曾被严格尝试过，但最终还是失败了。 完全放弃，由于健身成本上升，导致技术用于产生 易位株，除了不能将抗病基因与染色体上的 断点。重要的是，基因工程和合成生物学的最新进展允许这些 历史遗留问题必须彻底解决。此外，易位介导的基因驱动系统是 阈值依赖，因此具有几个对社会和科学接受很重要的有吸引力的特征， 野生转基因释放：该系统是物种特异性的;物种之间的零水平传播;最小 与杀虫剂相反的生态影响;强大而牢不可破，与自私的无情联系在一起。 基因元件及其货物;如果需要，可以从野生群体中完全去除转基因。 因此，本项目将利用最前沿的应用合成生物学原理， 染色体易位在精确的位置在Ae。埃及伊蚊（Aim-1）。一旦携带易位的菌株 建立，这些将与野生遗传背景渗入，健身动态将被测量， 将进行小型实验室规模的驱动实验（Aim-2）。总的来说，一个成功的基于易位的 与疾病难治性基因相关的人口替代系统将对两者产生重大影响 人类健康以及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中设计，因此这种创新方法也可以在以后的昆虫中设计。 广泛的昆虫疾病媒介，彻底改变和现代化的昆虫种群控制领域。

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.