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.

寄生蜂在自然生态系统中数量众多且重要，并且通常被工业用于控制害虫。在这项工作中，PI将测试两种用于遗传修饰寄生蜂的新方法。一种方法依赖于将黄蜂暴露于转基因宿主，另一种方法依赖于向成年雌性黄蜂注射卵巢和胚胎特异性吸收的结构。这两种方法的成功都将彻底改变我们在寄生蜂中询问基因功能的能力。这些信息将有助于生物经济，因为它将提供操纵寄生蜂的手段，以利于控制害虫。 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导人。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。宿主-寄生虫相互作用，如捕食者-猎物相互作用，是自然界中最重要的生态相互作用，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。寄生蜂是多种多样的，它们感染大多数昆虫物种，并且它们是用于对抗农业害虫的最常见的生物防治剂之一。不幸的是，标准的功能基因组学方法，如RNAi和CRISPR，在寄生蜂中是不够的，因为它们必须在宿主体内完成幼年发育，因此不符合标准的注射程序。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黄蜂靶向dsRNA的普遍表达是否会导致黄蜂寄生虫中的系统性RNAi敲除。这种新方法将利用这样的事实，即可以喂食双链RNA以诱导RNAi，并且内寄生蜂通过消耗宿主组织获得大部分营养。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组分（Cas9/gRNA）被贩运到黄蜂卵巢的发育胚胎中，从而导致基因敲除。新方法依赖于注射到较老的自由生活的生命阶段（如黄蜂蛹），其中许多胚胎可以通过一次注射转化。该奖项由综合有机体系统部的共生、防御和自我认可以及通过基因组学工具进行发现项目共同资助。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。