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

DNA测序的最新技术进步推动了生物学许多领域的重要新发现。“基因组革命”提供了丰富的数据，揭开了各种生物如何进化、相互作用和对环境做出反应的奥秘。然而，一个主要的障碍是绝大多数研究都是相关的。允许研究人员精确定位基因和特征之间因果关系的工具有限。这项研究将应用最新技术来研究一种新兴的生物生物学模型--黄蜂--的基因功能。这些昆虫，俗称纸蜂，是研究得很好的世界性群居昆虫。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的生物生物学问题(包括物种形成、行为、发育和神经科学)。该项目将采用两项尖端基因操作技术，并集中精力在实验室培育和维护菌落，以测试与着色、学习、攻击性和发育等特征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，这些研究人员对利用群居昆虫来解决生物生物学、应用昆虫学和农业中的各种问题感兴趣。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。结果、资源和方法将通过研讨会、实验室交流和工作坊广泛传播给研究界。该项目将以三管齐下的方式开发褐飞虱黄蜂功能基因组学资源。首先，研究人员将使用RNA干扰(RNAi)对候选基因功能进行生态相关测试。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学条件来提高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，旨在缩短种群周期，诱导筑巢和产卵，为研究提供持续的昆虫供应。这些发展将使Polistes成为培育功能基因组学稳定品系的易于处理的模型。第三，使用CRISPR/Cas9的基因编辑，该团队将通过注射胚胎和针对色素沉着基因黄色来培育第一代(G0)黄蜂，目标是生产具有非致命性但容易观察到的表型效应的转基因马赛克。成功的突变体G0将在实验室中用于生产稳定的黄色转基因株系，然后转移到其他感兴趣的目标基因。这项工作可能会将Polistes黄蜂转变为生物生物学众多领域的功能性基因组模型系统。这项工作也是独一无二的，因为它将允许在实验室和现场在受控条件下测试基因功能，提供了解决基因功能如何依赖于环境环境的主要优势。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.