SBIR/STTR Phase I: A High Throughput Gene Discovery System in Arabidopsis using Geminivirus gene expression vectors

This Small Business Technology Transfer (STTR) Phase I Project proposes to develop a geminivirus-derived gene expression system, based on cabbage leaf curl virus, to test the effect of agronomically important genes in a transient, high throughput model. While the genomics revolution is a major step toward understanding gene function, the foremost challenge is correlating bioinformatics data with actual functional responses. Paradigm Genetics, Inc., currently uses the plant model species, Arbidopsis thaliana in a pipeline developed specifically to address the bottleneck between sequence and functional data. Arabidopsis plants genetically engineered to over or underexpress single genes are analyzed in developmental, biochemical and molecular platforms. While this pipeline has been successful in many ways, two disadvantages remain : (1) the amount of time required to generate stable Arabidopsis transformants, and (2) the ability to test only one gene at a time. To address these issues, this proposal initiates the development of an additional platform using geminivirus vectors. First, a genetic screen of Arabidopsis ecotypes and a mutagenized population will be carried out. This screen is designed to isolate symptomless Arabidopsis lines supporting viral replication and spread. Second, based on the hypothesis that lower viral levels can decrease symptom development in the plant, the viral vector will be modified to replicate at lower levels. Results from this research will be used to start a high throughput assay to determine functions of agronomically important genes. Additionally, these results will lead to important agronomic understandings of virus/host interactions.