EAGER: Development Of Functional Genetic Tools For Endoparasitoid Wasps

寄生蜂数量众多，在自然生态系统中发挥着重要作用，也常被工业用来防治害虫。在这项工作中，pi将测试两种转基因寄生蜂的新方法。一种方法是将黄蜂暴露在转基因宿主中，另一种方法是向成年雌性黄蜂注射专门由其卵巢和胚胎吸收的结构物。这两种方法中的任何一种的成功都将彻底改变我们对寄生蜂基因功能的研究能力。这些信息将有助于生物经济，因为它将为控制害虫提供操纵寄生蜂的手段。该奖项还资助一名研究生的培训。因此，这笔资金正在培养与生物经济相关领域的下一代科学领袖。pi将通过研讨会、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。宿主-寄生虫的相互作用，就像捕食者-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，寄生蜂和它们的节肢动物宿主是这种关系最普遍的例子之一。寄生蜂种类繁多，能感染大多数昆虫，是农业害虫防治中最常用的生物防治剂之一。不幸的是，标准的功能基因组学方法，如RNAi和CRISPR，对寄生蜂来说是不够的，因为寄生蜂在寄主体内有义务完成幼体发育，因此对标准的注射程序是不适应的。为了充分利用寄生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在寄生蜂中的传递效率。在目标1中，pi将测试转基因宿主蝇中普遍表达的黄蜂靶向dsRNAs是否会导致黄蜂寄生虫的系统性RNAi敲低。这种新方法将利用膜翅目昆虫可以被喂食dsrna来诱导RNAi，以及类内寄生虫通过消耗宿主组织获得大部分营养物质的事实。在目标2中，pi将测试使用黄蜂特异性糖脂蛋白先导序列是否会导致CRISPR成分（Cas9/gRNA）被转运到黄蜂卵巢发育中的胚胎中导致基因敲除。新方法依赖于注射到较老的自由生活的生命阶段（如黄蜂蛹），在那里许多胚胎可以通过一次注射而转化。该奖项由综合有机体系统部门的共生、防御和自我识别以及通过基因组工具项目实现发现共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.