EDGE: Functional genomics in Polistes wasps, a model system in integrative organismal biology

EDGE：马蜂的功能基因组学，综合生物生物学的模型系统

基本信息

批准号：
1827567
负责人：
Amy Toth
金额：
$ 81.56万
依托单位：
Iowa State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1827567&HistoricalAwards=false
关键词：
EDGE Functional genomics Polistes wasps

项目摘要

Recent technological advances in DNA sequencing have fueled important new discoveries in many fields of biology. The "genomic revolution" has provided a wealth of data to unravel the mysteries of how diverse organisms evolve, interact, and respond to their environments. However, a major hurdle has been that the vast majority of studies are correlative. Tools allowing researchers to pinpoint causal associations between genes and traits are limited. This research will apply the latest technology to study gene function in an emerging model for organismal biology, Polistes wasps. These insects, commonly known as paper wasps, are well-studied, cosmopolitan, social insects. Because of their abundance in human environments, ease of experimental manipulation, and rapidly developing genomic resources, they are poised to address many integrative organismal biology questions (including speciation, behavior, development, and neuroscience). The project will employ two cutting edge technologies for gene manipulation along with focused efforts to breed and maintain colonies in the laboratory, in order to test causality for several genes associated with traits such as coloration, learning, aggression, and development. This work can have a catalytic impact on a growing community of researchers interested in using social insects to address a wide variety of questions in organismal biology, applied entomology and agriculture. Training and mentoring in the latest methodologies will be provided to one postdoctoral researcher, one graduate student, one research technician, and numerous undergraduates. Results, resources and methods will be disseminated widely to the research community through symposia, lab exchanges, and workshops.This project will develop resources for functional genomics in Polistes fuscatus wasps using a three-pronged approach. First, the researchers will use RNA-interference (RNAi) for ecologically relevant tests of candidate gene function. Functional genomic tests can be performed in the wild, allowing for assessment of gene function in an ecologically relevant context. The project will improve RNAi efficiency through experimenting with technical parameters and biological conditions and test candidate genes for roles in phenotypes such as caste development and cognition. Second, the researchers will establish methods for controlled mating in the laboratory, and continuous rearing, aiming to contract the colony cycle, induce nest founding, and induce egg laying to provide continuous availability of insects for research. These developments will establish Polistes as a tractable model for rearing stable lines for functional genomics. Third, using gene editing with CRISPR/Cas9, the team will produce first generation (G0) wasps by injecting embryos and targeting the pigmentation gene yellow, with the goal of producing transgenic mosaics with non-lethal but readily observable phenotypic effects. Successful mutant G0s will be used to produce stable transgenic lines in the lab for yellow and then move on to other target genes of interest. This work may transform Polistes wasps into a functional genomic model system for numerous areas of organismal biology. The work is also unique in that it will allow for tests of gene function under controlled conditions in the laboratory as well as in the field, providing the major advantage of addressing how gene function depends on environmental context.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.

DNA测序的最新技术进步推动了许多生物学领域的重要新发现。 “基因组革命”提供了大量数据，以揭示各种有机体如何发展，互动和对其环境做出反应的奥秘。但是，一个主要的障碍是绝大多数研究都是相关的。允许研究人员确定基因与特征之间的因果关系的工具受到限制。这项研究将应用最新技术来研究有机生物学新兴模型Polistes Wasps的基因功能。这些昆虫通常被称为纸黄蜂，是经过充分研究的，是国际化的社会昆虫。由于它们在人类环境中的丰富性，易于实验操作以及快速发展的基因组资源，因此他们有望解决许多综合生物学问题（包括物种，行为，发展和神经科学）。该项目将采用两种尖端技术来进行基因操纵，并重点努力繁殖和维持实验室的菌落，以测试与色彩，学习，侵略和发展等特征相关的几种基因的因果关系。这项工作可能会对有兴趣使用社交昆虫来解决生物学，应用昆虫学和农业的各种问题的越来越多的研究人员产生催化影响。将向一位博士后研究员，一名研究生，一名研究技术员和众多本科生提供最新方法的培训和指导。结果，资源和方法将通过研讨会，实验室交流和讲习班广泛地传播到研究界。本项目将使用三方面的方法来开发Polistes Fuscatus Wasp的功能基因组学资源。首先，研究人员将使用RNA干扰（RNAI）进行候选基因功能的生态相关测试。可以在野外进行功能基因组检验，从而可以在生态相关的环境中评估基因功能。该项目将通过实验技术参数和生物条件，并测试候选基因在种姓发展和认知等表型中的作用来提高RNAi效率。其次，研究人员将建立实验室中控制交配的方法，并持续饲养，旨在收缩菌落周期，引起巢穴的成立，并诱导卵子铺设以提供昆虫的连续可用性进行研究。这些发展将建立Polistes作为用于培养功能基因组学稳定线路的可进行的模型。第三，使用CRISPR/CAS9的基因编辑，该团队将通过注射胚胎和靶向色素沉着基因黄色来产生第一代黄蜂（G0）黄蜂，其目的是产生具有非杀伤性但易于观察到的表型效应的转基因镶嵌物。成功的突变体G0将用于在实验室中产生稳定的转基因线，以换黄色，然后转向其他感兴趣的目标基因。这项工作可能会将polistes黄蜂转变为一个有机生物学领域的功能基因组模型系统。这项工作也是独一无二的，因为它将允许在实验室和现场受控条件下对基因功能进行测试，从而提供了解决基因功能如何取决于环境环境的主要优势。该奖项反映了NSF的法定任务，并认为通过基金会的知识和更广泛的影响，可以通过评估来审查Criteria通过评估来进行评估。