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

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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.