As the scientific community progresses toward the completion of genome sequences, the availability of genome-scale tools for investigating gene function becomes critical. Of course, conventional mutagenesis and genetics will always be a mainstay of such efforts; however, more directed tools that selectively attack the expression of genes in a targeted fashion would accelerate reverse genetic approaches to understanding the function of genes and gene families. With the discovery of RNA interference (RNAi), the design of such tools has become possible, and genome-scale projects are already well underway in worm, Drosophila and human systems.
In this project, we will lay the groundwork for rapid generation of genome-scale resources for trigging post-transcriptional gene silencing of plant genes. Initially, we will use tobacco and Arabidopsis and once we validate our approach in these models, we will move on toward application of these technologies in rice. Flexible vectors will be developed that incorporate short and long hairpin RNAs and their efficacy will be tested in plant cell cultures and intact plants. The optimized vectors will be made available as well as silenced plants. The long-term goal is to use these vectors to develop genome scale RNAi libraries for plants in a rapid and cost-effective way.
This work is appropriate for support through a Small Grant for Exploratory Research for several reasons. Not only does it represent preliminary work on a risky and untested idea but success in this venture would also be transforming for the plant community in that it would provide the basis for building new and widely applicable tools. Furthermore, success would likely provide a new technology that would catalyze rapid advances in a broad spectrum of areas in plant biology.
Project outcomes Vectors will be distributed through Open Biosystems (www.openbiosystems.com) and plant materials will be available from Cold Spring Harbor Laboratory.
