Self-eliminating strategy to control gene drive

控制基因驱动的自我消除策略

• 批准号: 10202464
• 负责人: Zach N. Adelman
• 金额: $ 74.19万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-25 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10202464
• 关键词: Aedes; Alleles; Animal Model; Arboviruses; Arthropods; Behavior; Biological Models; Biology; CRISPR gene drive; CRISPR/Cas technology; Chikungunya virus; Communities; Culicidae; DNA; DNA Repair; Data; Dengue; Development; Direct Repeats; Disease Vectors; Drosophila genus; Drosophila melanogaster; Ecosystem; Effectiveness; Environment; Eukaryota; Excision; Funding; Future; Genes; Genetic Models; Health; Human; Infrastructure; International; Intervention; Length; Longevity; Malaria; Measures; Modeling; Mosquito Control; Natural Resistance; Nature; Population; Population Sizes; Process; Resistance; Series; Site; System; Tail; Target Populations; Technology; Testing; Time; Transgenes; Transgenic Organisms; Yellow Fever; base; combat; density; ds-DNA; expectation; fly; gene drive system; genetic element; hazard; innovation; malaria transmission; novel; nuclease; pathogen; repaired; resistance gene; restoration; safety testing; simulation; targeted nucleases; trait; vector; vector mosquito

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.

长期以来，将病原体抗性基因传递给蚊子种群的能力 寻求作为阻断登革热或疟疾传播的潜在替代方案，其中资金和 基础设施是有效控制蚊子的限制因素。近期的发展 基于CRISPR/Cas9的有效基因驱动转基因，很大程度上解决了技术难题 实现超孟德尔基因渗入面临着挑战，但目前还没有办法 一旦释放，就控制或召回此类遗传元件，并在相关部门进行安全测试 环境有问题。果蝇是一种极其可追踪的遗传模型 埃及伊蚊是登革热、黄热病和基孔肯雅热的主要传播媒介 病毒，以及用于研究其他蚊子的模型系统。在这个项目中，我们将 使用黑腹果蝇和埃及伊蚊来评估转基因自我消除策略 转基因可以被预先编程，首先将自身驱动到群体中，然后 在没有实验者任何干预的情况下将其自身从群体中移除。我们将 表征自消除系统的一些限制参数，例如直接 重复长度（目标 1）以及核酸酶靶向位点的数量和类型（目标 2）。最后，我们将 在果蝇和果蝇的活跃基因驱动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 在潜在的现场发布之后。因此，我们预计该项目将极大地改变 关于整个基因驱动技术的国内和国际对话，并将 提高对未来任何试验中可能产生病原体抗药性的期望 蚊子。