IOS EDGE: Accelerating arthropod genetic manipulation through ReMOT Control

非技术性段落：基因操作是解决节肢动物研究问题的有力技术。目前的方法依赖于通过胚胎显微注射（EM）将基因编辑材料传递到节肢动物卵子。然而，EM非常具有挑战性，仅限于少数节肢动物物种，即使在优化物种中也是低效的。目前迫切需要开发一种简单的节肢动物遗传操作方法，这种方法对许多研究人员来说都是容易获得的，并且通常适用于大量节肢动物物种。我们已经开发了一种称为受体介导的卵巢货物转导或远程控制的技术，通过在卵子发育期间将基因编辑货物轻松注射到雌性节肢动物体内，专门将基因编辑货物传递到发育中的节肢动物种系。远程控制可以将基因改造技术的力量带给任何模型或非模型物种，而无需注射胚胎，允许任何实验室使用这些强大的工具来解决他们的研究问题。通过研讨会、社交媒体、专题讨论会和公开试剂，我们将确保任何感兴趣的研究人员都可以使用远程控制技术。我们还将利用许多外展场所向公众宣传这些神奇工具的好处。我们将开展外展活动，并利用现有的外展场所，吸引K-12学生和公众成员，教育他们这些技术如何有益于他们的日常生活。技术段落：远程控制技术是基于对发展中的节肢动物种系具有非常特异性的小肽配体的鉴定。这些配体用于将CRISPR/Cas9核糖核蛋白复合物在注入卵黄变性雌性后转导到发育中的卵子。在本提案中，我们将开发跨不同节肢动物类群的特定和通用远程控制技术，向所有对基因工程技术感兴趣的研究人员开放该技术的真正力量。远程控制将打破基因改造的障碍，使不同动物系统的研究人员能够超越相关性，精确地研究基因功能。我们这一提议的总体概念目标不亚于所有研究节肢动物系统的研究人员的基因编辑能力的完全民主化，无论是模型还是非模型。这一目标将通过以下三个具体目标来实现：1)确定物种特异性和通用的配体，将远程控制技术应用于多种卵生和胎生节肢动物物种；2)利用鉴定的配体将CRISPR/Cas9转导至宿主种系，在不同节肢动物物种中进行靶向基因缺失和基因敲入；3)向参与动物行为、动物生理学、昆虫-植物相互作用、可持续农业和公共卫生研究的科学家广泛传播远程控制信息、技术和方法。在研究项目结束时，这种变革性技术将使基因改造技术进入日常科学家的范围，无论他们的研究系统如何。

Non-technical paragraph: Genetic manipulation is a powerful technique for addressing research questions in arthropods. Current approaches rely upon delivering gene-editing material to arthropod eggs by embryonic microinjection (EM). However, EM is very challenging, is limited to a small number of arthropod species, and is inefficient even in optimized species. There is a critical need to develop methods for arthropod genetic manipulation that are simple, accessible for many researchers and generally compatible for a large variety of arthropod species. We have developed a technology called Receptor-Mediated Ovary Transduction of Cargo, or ReMOT Control, to specifically deliver gene-editing cargo to the developing arthropod germline by easy injection into female arthropods during egg development. ReMOT Control can bring the power of genetic modification technology to any model or non-model species without the need for injecting embryos, allowing any lab to use these powerful tools for their research questions. Using workshops, social media, symposia, and by making reagents publically available, we will ensure that the ReMOT Control technology is available to any interested researchers. We will also use numerous outreach venues to educate the public about the benefits of these amazing tools. We will develop outreach activities and leverage existing outreach venues that engage K-12 students and members of the public to educate them about how these techniques can benefit their everyday lives. Technical paragraph: The ReMOT Control technology is based on the identification of small peptide ligands with very specific tropism for the developing arthropod germline. These ligands are used to transduce the CRISPR/Cas9 ribonucleoprotein complex to developing eggs after injection into vitellogenic females. In this proposal, we will develop both specific and generalizable ReMOT Control technologies that work across diverse arthropod taxa, opening the true power of this technique to all researchers interested in genetic engineering techniques. ReMOT Control will break down barriers to genetic modification, allowing researchers in diverse animal systems to move beyond correlation to accurately and precisely study gene function. Our overarching conceptual goal for this proposal is nothing less than the complete democratization of gene editing capability for all researchers working in any arthropod system, be it model or non-model. This goal will be realized by the following three Specific Aims: 1) Identify species-specific and universal ligands for applying ReMOT Control technology to diverse oviparous and viviparous arthropod species; 2) Use identified ligands to transduce CRISPR/Cas9 to the host germline for targeted gene deletion and gene knock-in in diverse arthropod species; 3) Disseminate ReMOT Control information, technology and methodology to broad communities of scientists involved in research on animal behavior, animal physiology, insect-plant interactions, sustainable agriculture, and public health. By the end of the research project this transformative technology will bring genetic modification technology within the reach of everyday scientists regardless of their research system.