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EDGE FGT: Bee Functional Genomics Using Engineered Symbionts

EDGE FGT：使用工程共生体的蜜蜂功能基因组学

基本信息

批准号：
2103208
负责人：
Jeffrey Barrick
金额：
$ 182万
依托单位：
University of Texas at Austin
依托单位国家：
美国
项目类别：
Continuing Grant
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-15 至 2025-07-31
项目状态：
未结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2103208&HistoricalAwards=false
关键词：
EDGE FGT Bee Functional Genomics

项目摘要

Insects are among the most widespread and diverse animals on our planet. They have critical roles in natural ecosystems and agriculture and have evolved unique biomaterials and lifestyles. Scientific tools for studying the functions of the genes responsible for these traits are well-established for only a few types of insects, such as fruit flies. Genetic tools do not exist for millions of other species. Honey bees and bumblebees are economically important as widespread pollinators of food crops and are scientifically interesting due to the complex social behaviors observed in bee colonies. Currently, there are few effective tools for studying the functions of bee genes. This project will develop and disseminate a toolkit that allows researchers to alter the expression of bee genes by engineering their native symbiotic gut bacteria. This technology will enable studies of how specific genes contribute to bee physiology, development, and behavior. This work will contribute to understanding bee ecology and health in ways that are expected to benefit biodiversity and US food security in the long term. The technology for engineering symbiotic bacteria is expected to be widely applicable to studying other insect species. These research and outreach activities will be integrated with education by supporting two experiential learning courses that are part of the Freshman Research Initiative program at The University of Texas at Austin. Supporting this program will foster the development of a diverse science and technology workforce by involving underrepresented and first-generation college students in genuine research experiences.Silencing the expression of a gene by inducing an RNA interference (RNAi) response is a common approach for performing studies of gene function in invertebrates. However, delivering enough double-stranded RNA to achieve sufficient gene knockdown through injection or feeding is expensive and ineffective in many insects, including bees. In this project, a FUnctional Genomics Using Engineered Symbionts (FUGUES) methodology will be developed and applied to honey bees (Apis mellifera) and bumblebees (Bombus spp.). In FUGUES, microbial symbionts are engineered to continuously produce and deliver double-stranded RNA to induce a targeted RNAi response in their host. Newly emerged bees colonized with an engineered bacterial symbiont exhibit reduced expression of a target gene throughout the bee body, enabling one to ascertain the function of a bee gene and its role in determining specific phenotypes. There are key advantages of using FUGUES to study gene function over current techniques that generate transgenic animals: it can be accomplished more quickly, it can be conducted in high-throughput when coupled with insect colonization via feeding, and it can be applied to species, such as bees, with mating systems and collective behaviors that complicate using genome editing techniques. Improving and disseminating the FUGUES tools created in this work will broadly enable studies of genes underlying bee behavior, development, and physiology. These tools will likely also be useful for studying many other insect species and other organisms that harbor symbiotic bacteria.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.

昆虫是地球上分布最广泛、种类最丰富的动物之一。它们在自然生态系统和农业中发挥着关键作用，并进化出了独特的生物材料和生活方式。研究负责这些性状的基因功能的科学工具仅适用于少数类型的昆虫，例如果蝇。对于数百万其他物种来说，遗传工具并不存在。蜜蜂和熊蜂作为广泛的粮食作物授粉者，在经济上很重要，并且由于在蜂群中观察到的复杂的社会行为而在科学上很有趣。目前，研究蜜蜂基因功能的有效工具很少。该项目将开发并传播一个工具包，使研究人员能够通过改造蜜蜂的天然共生肠道细菌来改变蜜蜂基因的表达。这项技术将有助于研究特定基因如何影响蜜蜂的生理、发育和行为。这项工作将有助于了解蜜蜂生态和健康，从长远来看，有望有利于生物多样性和美国粮食安全。共生细菌工程技术有望广泛应用于其他昆虫物种的研究。这些研究和外展活动将通过支持两门体验式学习课程与教育相结合，这门课程是德克萨斯大学奥斯汀分校新生研究计划的一部分。支持该计划将通过让代表性不足的第一代大学生参与真正的研究经验来促进多元化科学和技术劳动力的发展。通过诱导 RNA 干扰 (RNAi) 反应来沉默基因的表达是进行无脊椎动物基因功能研究的常见方法。然而，通过注射或喂食输送足够的双链RNA以实现足够的基因敲除是昂贵的，并且对包括蜜蜂在内的许多昆虫来说是无效的。在该项目中，将开发一种使用工程共生体 (FUGUES) 的功能基因组学方法，并将其应用于蜜蜂 (Apis mellifera) 和熊蜂 (Bombus spp.)。在 FUGUES 中，微生物共生体被设计为连续产生和传递双链 RNA，以在宿主中诱导靶向 RNAi 反应。被工程化细菌共生体定植的新出现的蜜蜂在整个蜂体内表现出目标基因的表达减少，从而使人们能够确定蜜蜂基因的功能及其在确定特定表型中的作用。与目前产生转基因动物的技术相比，使用FUGUES研究基因功能具有关键优势：它可以更快地完成，当与通过喂养的昆虫定植相结合时可以以高通量进行，并且它可以应用于蜜蜂等物种，其交配系统和集体行为使使用基因组编辑技术变得复杂。改进和传播本工作中创建的 FUGUES 工具将广泛促进蜜蜂行为、发育和生理学基础基因的研究。这些工具也可能有助于研究许多其他昆虫物种和其他含有共生细菌的生物体。该奖项反映了 NSF 的法定使命，并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，被认为值得支持。