Self-eliminating strategy to control gene drive

长期以来，向蚊子种群传递病原体抗性基因的能力 寻求作为阻断登革热或疟疾传播的潜在替代方案，在那里，资金和 基础设施是有效控制蚊子的限制因素。的最新发展 基于CRISPR/Cas9的有效基因驱动转基因在很大程度上解决了 实现超孟德尔渐近的挑战，然而，没有办法 控制或召回这些基因成分，一旦释放，在相关的 环境问题。果蝇是一种非常可追踪的遗传模型 生物体，埃及伊蚊是登革热、黄热病和基孔肯雅热的主要媒介 病毒，以及研究其他蚊子的模型系统。在这个项目中，我们将 同时使用黑腹木霉和埃及伊蚊来评估转基因自我消除策略 因此，转基因可以被预先编程，首先将自己送入种群，然后 在没有实验者任何干预的情况下将自己从种群中移除。我们会 描述自消除系统的一些限制参数，如直接 重复长度(目标1)和核酸酶靶点的数量和类型(目标2)。最后，我们会 在果蝇和果蝇的活跃基因驱动的背景下测试这种自我消除策略 蚊子(目标3)。我们的创新方法利用了自然发生的过程 在整个真核生物中都是保守的，以完全消除所有转基因序列 在潜在的野外释放之后。因此，我们预计这个项目将极大地改变 关于基因驱动技术的国内和国际对话，并将 提高对未来试验中产生病原体抗药性的可能性的期望 蚊子。

The ability to deliver pathogen-resistance genes into mosquito populations has long been sought as a potential alternative for disrupting dengue or malaria transmission where funds and infrastructure are the limiting factors in effective mosquito control. The recent development of effective gene drive transgenes based on CRISPR/Cas9 has largely solved the technical challenges of achieving super-Mendelian introgression, however there exists no means to control or recall such genetic elements once released making safety testing in the relevant environments problematic. Drosophila melanogaster is an extremely trackable genetic model organism, while Aedes aegypti is the main vector of dengue, yellow fever and chikungunya viruses, as well as a model system for studies of other mosquitoes. In this project, we will employ both D. melanogaster and A. aegypti to evaluate a transgene self-elimination strategy whereby a transgene can be pre-programmed to first drive itself into a population and then remove itself from the population without any intervention from the experimenter. We will characterize some of the limiting parameters of the self-elimination system, such as direct repeat length (Aim 1) and number and type of nuclease targeting sites (Aim 2). Finally, we will test this self-elimination strategy in the context of an active gene drive in both flies and mosquitoes (Aim 3). Our innovative approach takes advantage of naturally occurring processes that are conserved throughout eukaryota to completely eliminate all transgenic sequences following potential field releases. Thus, we anticipate that this project will dramatically alter the National and International conversations concerning gene drive technology as a whole, and will raise expectations for what is possible in any future trial to generate pathogen-resistant mosquitoes.