SGER: Developing Methods for Global Expression Analysis at Cellular Resolution

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0127676BenfeyA critical first step in characterizing gene function is defining the expression pattern of a gene. A novel methodology is being developed in plants to rapidly determine the cell-type specific expression of thousands of transcripts in concert. The technique involves separating plant cells by protoplasting transgenic lines that express GFP in specific cell types, obtaining an enriched population of GFP expressing cells using a Fluorescence Activated Cell Sorter (FACS), and applying mRNA isolated from sorted cells to microarrays. The result is an expression profile from a specific cell type. Extensive controls have been performed and a first round of the entire protocol with a single transgenic line has been completed. The results are extremely promising with microarray profiles of specific genes in agreement with their independently confirmed expression patterns in the Arabidopsis root. The aims of this proposal are to: 1) Further test and develop the methodology. 2) Expand the number of sorted cell lines. 3) Develop computational tools to analyze and disseminate the data. The risk associated with this project is balanced by the enormous potential benefit to the plant community.