"SGER: Development of Tools for High Throughput RNA Interference in Plants."

随着科学界朝着完成基因组序列的方向发展，研究基因功能的基因组规模工具的可用性变得至关重要。当然，传统的诱变和遗传学将永远是这类努力的支柱；然而，更直接的工具，有针对性地攻击基因的表达，将加速反向遗传方法来理解基因和基因家族的功能。随着RNA干扰（RNAi）的发现，设计这样的工具已经成为可能，基因组规模的项目已经在蠕虫、果蝇和人类系统中顺利进行。在本项目中，我们将为快速生成触发植物基因转录后基因沉默的基因组级资源奠定基础。最初，我们将使用烟草和拟南芥，一旦我们在这些模型中验证了我们的方法，我们将继续将这些技术应用于水稻。将开发包含短发夹rna和长发夹rna的柔性载体，并在植物细胞培养和完整植物中测试其功效。将提供优化的载体以及沉默的植物。长期目标是利用这些载体快速、经济地开发植物基因组规模的RNAi文库。由于几个原因，这项工作适合通过探索性研究小额赠款获得支持。它不仅代表了对一个有风险和未经检验的想法的初步工作，而且这次冒险的成功也将改变植物界，因为它将为建立新的和广泛适用的工具提供基础。此外，成功可能会提供一种新技术，促进植物生物学广泛领域的快速发展。项目成果载体将通过Open Biosystems （www.openbiosystems.com）发布，植物材料将从冷泉港实验室获得。

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 outcomesVectors will be distributed through Open Biosystems (www.openbiosystems.com) and plant materials will be available from Cold Spring Harbor Laboratory.