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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”提供了丰富的数据来揭开各种生物如何进化、相互作用和对环境作出反应的奥秘。然而，一个主要的障碍是，绝大多数研究都是相关的。让研究人员能够确定基因和性状之间因果关系的工具是有限的。这项研究将应用最新技术来研究生物生物学新兴模型Polistes wasps中的基因功能。这些昆虫，通常被称为纸黄蜂，是一种被广泛研究的世界性社会昆虫。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，它们准备解决许多综合生物学问题（包括物种形成，行为，发育和神经科学）。该项目将采用两种尖端技术进行基因操作，沿着集中精力在实验室中繁殖和维持菌落，以测试与颜色，学习，侵略和发育等性状相关的几个基因的因果关系。这项工作可以对越来越多的研究人员产生催化作用，这些研究人员有兴趣利用社会昆虫来解决有机体生物学，应用昆虫学和农业中的各种问题。将向一名博士后研究员、一名研究生、一名研究技术员和许多本科生提供最新方法的培训和指导。研究结果、资源和方法将通过研讨会、实验室交流和讲习班广泛传播给研究界。本项目将使用三管齐下的方法开发用于褐蜂功能基因组学的资源。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。功能基因组测试可以在野外进行，允许在生态相关的背景下评估基因功能。该项目将通过试验技术参数和生物条件来提高RNAi效率，并测试候选基因在种姓发育和认知等表型中的作用。其次，研究人员将建立实验室控制交配和连续饲养的方法，旨在缩短殖民地周期，诱导筑巢，诱导产卵，为研究提供持续的昆虫。这些发展将建立Polistes作为一个听话的模式，饲养稳定的功能基因组学系。第三，使用CRISPR/Cas9进行基因编辑，该团队将通过注射胚胎并靶向色素沉着基因黄色来产生第一代（G 0）黄蜂，目标是产生具有非致命但易于观察的表型效应的转基因嵌合体。成功的突变体G 0将用于在实验室中产生稳定的黄色转基因株系，然后转移到其他感兴趣的靶基因。这项工作可能会将Polistes黄蜂转化为生物生物学许多领域的功能基因组模型系统。这项工作的独特之处还在于，它将允许在实验室和野外的受控条件下测试基因功能，提供了解决基因功能如何取决于环境背景的主要优势。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。