A Resource Center for Tetrahymena Thermophila

项目总结/摘要 该提案寻求继续为四膜虫股票中心提供资金，以维持其目前的 它将继续加强其业务，扩大其能力，并确保其作为整个社区的资源的可持续性。一个关键 作为真核细胞和分子生物学的模型，嗜热四膜虫一直是我们研究的工具。 了解与人类健康和疾病直接相关的广泛生物现象 包括癌症、不育、衰老、呼吸和神经内分泌功能障碍。四膜虫还显示 作为生产重组蛋白的平台，包括疫苗抗原和困难的， 表达人类离子通道，并作为一个有用的教学工具，在K12和本科课堂。 具体来说，我们对资源的目标是：1）继续作为一个接受新菌株的菌株库， 将T.以合理的价格向感兴趣的用户提供嗜热菌; 2）完成注释 目前存放在中心的档案菌株，并获得下一代克隆载体的微和 大核基因组编辑; 3）提供扩展的服务并开发创新的营销方法， 增加收入。此外，为了确保资源的长期稳定，我们计划重新安置 将存储库从康奈尔大学迁移到圣路易斯的华盛顿大学; 5）将所有数据从遗留数据库迁移 四膜虫基因组数据库（TGD）模式到CHADO，并将其网站托管功能从布拉德利转移到CHADO 美国国家科学基金会资助的国家基因组分析支持中心（NCGAS）在印第安纳州大学;和， 6)重新设计TGD网站，通过Tripal框架显示相关数据。我们的其他研究目标 7）开发缺乏功能性核糖体DNA位点的遗传菌株，以实现无选择创造 新的细胞系;以及，8）开发CRISPR介导的基因组编辑工具，以增强CRISPR的整体效用。 四膜虫平台。

Project Summary / Abstract This proposal seeks continued funding for the Tetrahymena Stock Center in order to maintain its current operations, expand its capabilities and ensure its sustainability as a resource for the community at-large. A key model for eukaryotic cell and molecular biology, Tetrahymena thermophila has been instrumental to our understanding of a wide range of biological phenomena with direct relevance to human health and disease including cancer, infertility, aging, respiratory and neuroendocrine dysfunction. Tetrahymena has also shown great promise as a platform for the production of recombinant proteins, including vaccine antigens and difficult- to-express human ion channels, and serves as a useful teaching tool in K12 and undergraduate classrooms. Specifically, our aims for the Resource are to 1) continue to act as a strain repository accepting new strains and making live cultures of T. thermophila available to interested users at reasonable cost; 2) complete the annotation of archival strains currently housed at the Center and acquire next-generation cloning vectors for micro- and macronuclear genome-editing; 3) offer expanded services and develop innovative marketing approaches to increase revenues. Additionally, to ensure insure long-term stability of the resource we plan to 4) relocate the repository from Cornell University to Washington University in St. Louis; 5) migrate all data from the legacy Tetrahymena Genome Database (TGD) schema to CHADO and transfer its web-hosting functions from Bradley University to the NSF-funded National Center for Genome Analysis Support (NCGAS) at Indiana University; and, 6) redesign the TGD website to display relevant data via the Tripal framework. Our additional Research aims are to: 7) develop genetic strains lacking a functional ribosomal DNA locus to enable selection-free creation of new cell lines; and, 8) develop CRISPR-mediated genome editing tools to enhance the overall utility of the Tetrahymena platform.