TRPGR: Maize Cell Genomics: Resources for Visualizing Promoter Activity and Protein Dynamics using Fluorescent Protein Lines

PI：David Jackson(冷泉港实验室)合作PI：Anne Sylvester(怀俄明大学)Agnes Chan(J.Craig Venter Institute)智力价值。玉米是粮食作物功能基因组学的一个强大的模式系统，因为它的参考基因组最近已经被测序，功能工具也在增加。然而，需要新的方法来研究细胞和组织内的基因功能，以了解细胞和亚细胞水平上的蛋白质功能，这最终将为作物改良提供指导信息。该项目的目标是在玉米中产生工具，这些工具可以用来在一些特定的选定组织或细胞类型中驱动任何基因的表达。这些工具的使用将使报告基因以前所未有的空间和时间分辨率可视化，并将使功能基因过度表达或基因敲除研究成为可能。此外，该项目将利用新改进的荧光蛋白质颜色标签来标记亚细胞或发育隔间，并将开发用于种植玉米植株的实时成像技术，以演示所有报告系在功能研究中的用途。因此，该项目将通过提供一种手段来了解特定细胞和组织在发育过程中的基因功能，从而改变玉米的功能遗传学实验。该项目将向研究界提供稳定转化的种子的永久库存，用于50个组织特异性激活剂/报告品系、20个荧光标记的蛋白质品系，以及充分表征活体成像方法。它还将开发一条处理大型图像数据集的管道，该管道将适用于其他项目。所有资源，包括图像、种子和基因结构，都将通过项目网站http://maize.jcvi.org/cellgenomics.公开提供为了长期可用，图像将被迁移到社区数据库MaizeGDB，种子将被存放在玉米遗传资源中心。更广泛的影响。该项目将通过在小大角学院为美国原住民学生组织一年一度的遗传学研讨会，将教育和外联结合在一起。小大角学院是乌鸦之国的部落学院。该项目还将通过生成玉米细胞和生长植物的3D互动动画，展示亚细胞室和基因表达数据，将功能研究整合到教育工具中。这些动画将由三个目标群体进行Beta测试，包括小大角学院的美国原住民学生，加拉德特大学细胞生物学课上的聋人和重听学生，以及怀俄明大学的生物学学生。动画将公之于众，并纳入参与机构教授的课程课程。

PI: David Jackson (Cold Spring Harbor Laboratory)Co-PIs: Anne Sylvester (University of Wyoming)Agnes Chan (The J. Craig Venter Institute)Intellectual Merit. Maize is a powerful model system for functional genomics in the grain crops, because its reference genome has recently been sequenced and functional tools are increasing. However, new methods of investigating gene function within cells and tissues are needed to understand protein functions at the cellular and sub-cellular level, which will ultimately provide guiding information for crop improvement. This project aims to generate tools in maize that can be used to drive expression of any gene in a number of specific chosen tissue or cell types. The use of these tools will permit the visualization of reporter genes with unprecedented spatial and temporal resolution, and will also enable functional gene over-expression or gene knockout studies. In addition, this project will make use of newly improved fluorescent protein color tags to label sub-cellular or developmental compartments, and will develop live imaging techniques for growing maize plants to demonstrate the utility of all of the reporter lines for functional studies. The project will therefore transform functional genetic experiments in maize, by providing a means to understand gene function in specific cells and tissues during development. The project will deliver to the research community a permanent stock of stably transformed seeds for 50 tissue specific activator/reporter lines, 20 fluorescent-tagged protein lines, as well as fully characterized in vivo imaging methods. It will also develop a pipeline for handling large image datasets that will be applicable to other projects. All resources, including images, seeds and gene constructs, will be publicly available via the project website http://maize.jcvi.org/cellgenomics. For long-term availability, images will be migrated to the community database, MaizeGDB, and seeds will be deposited with the Maize Genetics Stock Center. Broader Impacts. The project will integrate education and outreach by organizing an annual Genetics Workshop for Native American students at Little Big Horn College, a Tribal College to the Crow Nation. The project will also integrate functional research into educational tools by generating 3D interactive animations of a maize cell and growing plant, displaying sub-cellular compartments and gene expression data. The animations will be beta-tested by three target groups, including Native American students at Little Big Horn College, deaf and hard of hearing students in a cell biology class at Gallaudet University, and University of Wyoming biology students. Animations will be made publicly available and incorporated in course curricula taught at the participating institutions.