EDGE FGT: Development of fibroblasts and organoids as tools for functional genomics in turtles, applicable to other non-mammalian vertebrates

爱荷华州立大学的这个项目将为海龟开发基因组工具，以促进这些非模式分类群(和其他爬行动物)的使用，以研究基因组到表型组的映射、基因功能、发育、基因组结构、对气候变化的适应反应等。海龟是生态学、进化论和其他主题的新兴模型，但要完全实现这一承诺，还需要基因组工具。该项目将为科学界提供一个生物库和适用于海龟和其他爬行动物的工具。讲习班、专题讨论会和出版物将使这些资源得以迅速传播。将提供不同职业发展水平的培训(本科生、研究生、博士后和教师)，包括来自代表性不足群体的个人。将开展公众宣传活动，让爱荷华州科学中心的儿童和成年人了解海龟生物学和普通科学。功能基因组资源将在彩绘甲鱼成纤维细胞和类有机物中开发，包括基因上调(慢病毒载体或RNA注射)、基因敲除(RNAi)和靶向突变工具(CRISPR-CAS9)。这些工具将进一步使海龟能够作为模型系统，通过测试基因组到现象组的映射来阐明基因调控、脊椎动物发育的基础以及结构变异对基因组功能的影响。使用成纤维细胞和类有机物质也解决了实验中鸡蛋季节性的瓶颈，使用瞬时转基因的功能基因组学方法缓解了研究长寿生物体的一些挑战。开发成纤维细胞和有机体作为功能基因组学的新系统，将促进对海龟和其他爬行动物气候变化影响的研究。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。

This project at Iowa State University will develop genomic tools for turtles to advance the use of these non-model taxa (and other reptiles) to study genome-to-phenome mapping, gene function, development, genome architecture, adaptive responses to climate change, and more. Turtles are an emerging model for ecology, evolution, and other topics, but genomic tools are needed for that promise to be fully realized. This project will provide the scientific community with a biobank and tools applicable to turtles and other reptiles. Workshops, symposia, and publications will enable the rapid dissemination of these resources. Training will be provided across levels of career development (undergraduate, graduate, postdoctoral, and faculty), including individuals from underrepresented groups. Public outreach activities will be developed to engage children and adults at the Science Center of Iowa to understand turtle biology and general science. Functional genomic resources will be developed in painted turtle fibroblasts and organoids, including gene upregulation (lentiviral vectors or RNA injection), gene knockdown (RNAi), and targeted mutagenesis tools (CRISPR-Cas9). The tools will further enable turtles as model systems to illuminate gene regulation, the underpinnings of vertebrate development, and the effects of structural variation on genome function by enabling tests of genome-to-phenome mapping. Using fibroblasts and organoids also solves the bottleneck from the seasonality of eggs for experimentation and functional genomic approaches with transient transgenics mitigates some of the challenge of studying long-lived organisms. Developing fibroblasts and organoids as new systems for functional genomics will catalyze the study of climate change effects in turtles and other reptiles.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.