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（怀明大学）Agnes Chan （J. Craig Venter研究所）由于玉米的参考基因组的测序和功能工具的不断增加，玉米是一个强大的谷类作物功能基因组学模型系统。然而，研究细胞和组织内基因功能的新方法需要在细胞和亚细胞水平上了解蛋白质的功能，这将最终为作物改良提供指导信息。这个项目的目的是在玉米中产生工具，可以用来驱动在许多特定选择的组织或细胞类型中的任何基因的表达。这些工具的使用将允许以前所未有的空间和时间分辨率可视化报告基因，也将使功能性基因过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颜色标签来标记亚细胞或发育区室，并将开发用于种植玉米植物的实时成像技术，以证明所有报告系在功能研究中的实用性。因此，该项目将通过提供一种方法来了解发育过程中特定细胞和组织中的基因功能，从而改变玉米的功能基因实验。该项目将为研究界提供50个组织特异性激活/报告系、20个荧光标记蛋白系以及完全表征的体内成像方法的稳定转化种子的永久库存。它还将开发一个处理大型图像数据集的管道，这将适用于其他项目。所有资源，包括图像、种子和基因构建，将通过项目网站http://maize.jcvi.org/cellgenomics公开提供。为了获得长期可用性，图像将迁移到社区数据库MaizeGDB，种子将存放在玉米遗传库存中心。更广泛的影响。该项目将通过在小比格霍恩学院（Crow Nation的一所部落学院）为美洲原住民学生组织一年一度的遗传学研讨会，将教育和推广结合起来。该项目还将通过生成玉米细胞和生长植物的3D互动动画，显示亚细胞区室和基因表达数据，将功能研究整合到教育工具中。这些动画将在三个目标群体中进行beta测试，包括小比格霍恩学院的美国原住民学生，加劳德特大学细胞生物学课上的聋哑和重听学生，以及怀俄明大学生物学学生。动画将公开提供，并纳入参与机构的课程。

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.