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

SBIR/STTR 第一阶段：使用双生病毒基因表达载体的拟南芥高通量基因发现系统

• 批准号: 0215013
• 负责人: Susanne Kjemtrup
• 金额: $ 10万
• 依托单位: ICORIA INC
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2003-10-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0215013&HistoricalAwards=false
• 关键词: SBIR; STTR; Phase; Throughput; Gene

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 towardunderstanding gene function, the foremost challenge is correlating bioinformatics data with actualfunctional 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.The commercial applications of this project are in crop plant improvement.

这个小企业技术转让（STTR）第一阶段项目提出开发一个双生病毒衍生的基因表达系统，基于卷心菜曲叶病毒，以测试农艺学上重要的基因在瞬时，高通量模型的影响。虽然基因组学革命是理解基因功能的重要一步，但最重要的挑战是将生物信息学数据与实际功能反应相关联。Paradigm Genetics，Inc.，目前使用的植物模式物种，Arbidopsis thaliana在一个专门开发的管道，以解决序列和功能数据之间的瓶颈。在发育、生物化学和分子平台上分析了基因工程改造成过表达或低表达单个基因的拟南芥植物。虽然这种管道在许多方面都是成功的，但仍然存在两个缺点：（1）产生稳定的拟南芥转化体所需的时间量，以及（2）一次只能测试一个基因的能力。为了解决这些问题，该提案启动了使用双生病毒载体的额外平台的开发。首先，将进行拟南芥生态型的遗传筛选和诱变群体。该筛选设计用于分离支持病毒复制和传播的无病毒拟南芥品系。第二，基于较低的病毒水平可以减少植物中症状发展的假设，病毒载体将被修饰以在较低水平复制。这项研究的结果将用于启动高通量测定，以确定农学上重要基因的功能。此外，这些结果将导致重要的农学理解病毒/宿主相互作用。该项目的商业应用是在作物改良。