EAGER: Development Of Functional Genetic Tools For Endoparasitoid Wasps

EAGER：开发内寄生蜂的功能遗传工具

• 批准号: 2022235
• 负责人: Todd Schlenke
• 金额: $ 30万
• 依托单位: University of Arizona
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2022235&HistoricalAwards=false
• 关键词: EAGER; Development; Functional; Genetic; Tools

Parasitic wasps are numerous and important in natural ecosystems, and are also commonly used by industry to control pest insects. In this work, the PIs will test two novel approaches for genetically modifying parasitic wasps. One method relies on exposing wasps to genetically-modified hosts, and the other relies on injecting adult female wasps with constructs that will specifically be taken up by their ovaries and embryos. Success of either of these methods would revolutionize our ability to interrogate gene function in in parasitic wasps. This information would be helpful for the bioeconomy because it would provide the means to manipulate parasitic wasps to the benefit of controlling pest insects. This award also funds training of a graduate student. Thus, this funding is training the next generation of leaders in science in an area relevant to the bioeconomy. The PIs will broadly publicize and disseminate their research to academia and biotech companies via conference symposiums, lab workshops, and video protocols. Host-parasite interactions, like predator-prey interactions, are some of the most important ecological interactions in nature, and parasitic wasps and their arthropod hosts are one of the most ubiquitous examples of this type of relationship. Parasitic wasps are diverse, they infect most insect species, and they are one of the most common biocontrol agents used against agricultural pests. Unfortunately, standard functional genomics methods like RNAi and CRISPR are inadequate in parasitic wasps, given that they obligately complete their juvenile development inside their hosts and are thus recalcitrant to standard injection procedures. To fully harness the potential of parasitic wasps as a model system, the PIs will develop novel applications of RNAi and CRISPR to maximize the efficiency of dsRNA and Cas9-gRNA delivery into wasps. In Objective 1, the PIs will test whether ubiquitous expression of wasp-targeted dsRNAs in transgenic host flies will cause systemic RNAi knockdown in wasp parasites. This new method would take advantage of the facts that hymenopterans can be fed dsRNAs to induce RNAi, and that endoparasitoids obtain a majority of their nutrients by consuming host tissues. In Objective 2, the PIs will test whether use of a wasp-specific glycolipoprotein leader sequence will cause CRISPR components (Cas9/gRNA) to be trafficked into the developing embryos of wasp ovaries to cause gene knockouts. The new method relies on injection into older free-living life stages (like wasp pupae), where many embryos can be transformed via a single injection. This award was co-funded by the Symbiosis, Defense and Self Recognition and Enabling Discovery through GEnomic Tools programs in the Division of Integrative Organismal Systems.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

寄生黄蜂在自然生态系统中数量众多且重要，也常被工业用来控制害虫。在这项工作中，PI 将测试两种对寄生黄蜂进行基因改造的新方法。一种方法依赖于将黄蜂暴露于转基因宿主中，另一种方法依赖于向成年雌性黄蜂注射可被其卵巢和胚胎特异性吸收的结构。这些方法中任何一种的成功都将彻底改变我们研究寄生黄蜂基因功能的能力。这些信息将有助于生物经济，因为它将提供操纵寄生黄蜂以利于控制害虫的方法。 该奖项还资助研究生的培训。因此，这笔资金正在培训生物经济相关领域的下一代科学领导者。 PI 将通过会议研讨会、实验室研讨会和视频协议向学术界和生物技术公司广泛宣传和传播他们的研究成果。宿主与寄生虫的相互作用，就像捕食者与猎物的相互作用一样，是自然界中最重要的生态相互作用之一，寄生黄蜂及其节肢动物宿主是这种关系最普遍的例子之一。寄生黄蜂多种多样，它们感染大多数昆虫物种，并且是针对农业害虫最常见的生物防治剂之一。不幸的是，RNAi 和 CRISPR 等标准功能基因组学方法在寄生黄蜂中是不够的，因为它们必须在宿主体内完成幼年发育，因此不符合标准注射程序。为了充分利用寄生黄蜂作为模型系统的潜力，PI 将开发 RNAi 和 CRISPR 的新应用，以最大限度地提高 dsRNA 和 Cas9-gRNA 递送到黄蜂中的效率。在目标 1 中，PI 将测试转基因宿主果蝇中针对黄蜂的 dsRNA 的普遍表达是否会导致黄蜂寄生虫中的系统性 RNAi 敲低。这种新方法将利用以下事实：膜翅目动物可以通过喂食 dsRNA 来诱导 RNAi，并且内寄生物通过消耗宿主组织来获取大部分营养。在目标 2 中，PI 将测试使用黄蜂特异性糖脂蛋白前导序列是否会导致 CRISPR 组件 (Cas9/gRNA) 被贩运到黄蜂卵巢发育中的胚胎中，从而导致基因敲除。新方法依赖于注射到较老的自由生活阶段（如黄蜂蛹），其中许多胚胎可以通过单次注射进行转化。该奖项由综合有机系统部门的共生、防御和自我识别以及通过基因组工具实现发现项目共同资助。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力优点和技术进行评估，被认为值得支持。更广泛的影响审查标准。