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IOS EDGE: Accelerating arthropod genetic manipulation through ReMOT Control

IOS EDGE：通过 ReMOT Control 加速节肢动物基因操作

基本信息

批准号：
1645331
负责人：
Jason Rasgon
金额：
$ 250万
依托单位：
Pennsylvania State Univ University Park
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-01 至 2025-05-31
项目状态：
未结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1645331&HistoricalAwards=false
关键词：
IOS EDGE Accelerating arthropod genetic

项目摘要

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.

非技术性段落：基因操作是解决节肢动物研究问题的一种强有力的技术。目前的方法依赖于通过胚胎显微注射（EM）将基因编辑材料递送到节肢动物卵中。然而，EM是非常具有挑战性的，仅限于少数节肢动物物种，即使在优化的物种是低效的。迫切需要开发简单、许多研究人员容易获得并且通常与多种节肢动物物种相容的节肢动物遗传操作方法。我们开发了一种名为受体介导的卵巢转运货物（Receptor-Mediated Ovary Transduction of Cargo，简称ReMOT Control）的技术，通过在卵子发育期间轻松注射到雌性节肢动物中，专门将基因编辑货物递送到发育中的节肢动物种系。ReMOT Control可以为任何模式或非模式物种带来基因改造技术的力量，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、专题讨论会和提供试剂，我们将确保任何感兴趣的研究人员都可以使用ReMOT Control技术。我们还将利用众多的外展场所来教育公众这些令人惊叹的工具的好处。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，教育他们这些技术如何使他们的日常生活受益。技术段落：ReMOT控制技术是基于对发育中的节肢动物生殖系具有非常特异性向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射到卵黄发生雌性中后转移到发育中的卵中。在这项提案中，我们将开发适用于不同节肢动物类群的具体和可推广的ReMOT控制技术，向所有对基因工程技术感兴趣的研究人员开放这项技术的真正力量。ReMOT Control将打破转基因的障碍，使不同动物系统的研究人员能够超越相关性，准确和精确地研究基因功能。我们对这一提议的总体概念目标是让所有在任何节肢动物系统中工作的研究人员的基因编辑能力完全民主化，无论是模型还是非模型。该目标将通过以下三个具体目的来实现：1）鉴定用于将ReMOT控制技术应用于不同卵生和胎生节肢动物物种的物种特异性和通用配体; 2）使用鉴定的配体将CRISPR/Cas9引入宿主种系，用于在不同节肢动物物种中靶向基因缺失和基因敲入; 3）向参与动物行为、动物生理学、昆虫与植物相互作用、可持续农业和公共卫生研究的广大科学家社区传播ReMOT控制信息、技术和方法。到研究项目结束时，这项变革性的技术将使普通科学家能够接触到转基因技术，无论他们的研究系统如何。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing.

将CRISPR-CAS9核糖核蛋白的靶向递送到节肢动物卵巢中，以进行可遗传的种系基因编辑。

DOI：
10.1038/s41467-018-05425-9
发表时间：
2018-08-01
期刊：
Nature communications
影响因子：
16.6
作者：
Chaverra-Rodriguez D;Macias VM;Hughes GL;Pujhari S;Suzuki Y;Peterson DR;Kim D;McKeand S;Rasgon JL
通讯作者：
Rasgon JL

Actin bundles play a different role in shaping scales compared to bristles in the mosquito Aedes aegypti