A Resource Center for Tetrahymena Thermophila

项目摘要/摘要 这项提案寻求继续为四膜虫库存中心提供资金，以维持其现有的 行动，扩大其能力，并确保其作为整个社区的资源的可持续性。一把钥匙 真核细胞和分子生物学的模型，嗜热四膜虫对我们的 对与人类健康和疾病直接相关的广泛生物现象的了解 包括癌症、不孕不育、衰老、呼吸和神经内分泌功能障碍。四膜虫也显示出 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，在K12和本科生课堂上作为有用的教学工具。 具体地说，我们对该资源的目标是1)继续充当菌种存储库，接受新的菌种和 以合理的价格向感兴趣的用户提供嗜热支原体的活培养；2)完成注释 目前保存在该中心的存档菌株，并获得下一代克隆载体，用于微型和 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4)将 从康奈尔大学到圣路易斯的华盛顿大学的存储库；5)迁移旧版本中的所有数据 将四膜虫基因组数据库(TGD)模式转移到Chado，并从Bradley转移其网络托管功能 印第安纳大学国家科学基金会资助的国家基因组分析支持中心(NCGAS)； 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基因座的遗传菌株，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，以提高 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.