Arabidopsis 2010: A Comprehensive Resource for Analysis of Arabidopsis Gene Function

Pi：Gregory J.Hannon(冷泉港实验室)Copis：Robert A.Martin enssen和W.Richard McCombie(冷泉港实验室)确定模式植物拟南芥中所有基因的功能，将对我们对植物生物学的理解和我们提高重要农业植物物种所需性状的能力产生深远影响。确定基因功能的传统方法通常依赖于创建非功能基因的突变版本。这是通过改变基因本身，然后将一个不起作用的基因的两个拷贝放在一种植物中来实现的。这一策略适用于约30,000个拟南芥基因中的大多数，这些基因以单拷贝形式存在于基因组中。然而，大约有8000个拟南芥基因以密切相关的拷贝家族的形式存在，这些拷贝用传统方法很难协调失活。CSHL 2010项目的目标是开发能够确定构成这些多基因家族的基因的功能的资源。为了实现这一点，该项目将利用一条保守的生物途径，作为基因沉默的可编程引擎。这种途径被称为RNA干扰或RNAi。作为对双链RNA的反应，RNAi关闭了与沉默触发器具有相同序列的任何基因的表达。与传统的突变体不同，表达双链RNA的人工基因的一个副本足以让一个多基因家族的所有成员沉默。此外，双链RNA的产生可以通过一种方式进行调节，将基因的失活限制在特定的组织或发育时期。RNAi将针对存在于拟南芥多基因家族成员的8000个基因中的每一个基因进行触发。此外，还将产生针对发育早期必不可少的基因的RNAi触发器，因为在选定的组织中或以定时方式关闭这些基因的活性的能力可能会揭示它们的功能方面，而这些功能是传统突变无法阐明的。通过该项目开发的资源将补充现有的突变体库，因为常规工具不适合确定多基因家族中的基因和具有早期致死表型的基因的功能。如果没有这种互补的方法，到2010年了解拟南芥中所有基因的功能的目标将是不可能实现的。所有资源都将通过公共储存库--拟南芥生物资源中心和商业试剂分销网络--开放生物系统开发，供科学界随时使用。有关资源状况、整个项目以及如何获得材料的信息，可从项目网站http://2010.cshl.edu.获得。有关参与调查的人员及其研究计划的信息也可供查阅。关于外展和培训，该项目将通过多兰DNA学习中心利用冷泉港实验室的努力，将产生的植物资源纳入其课程。

PI: Gregory J. Hannon (Cold Spring Harbor Laboratories)CoPIs: Robert A. Martienssen and W. Richard McCombie (Cold Spring Harbor Laboratories)Determining the function of all the genes in the model plant, Arabidopsis thaliana, will have a profound impact on both our understanding of plant biology and our ability to enhance desirable traits in agriculturally important plant species. Conventional methods for determining gene function generally rely on creating mutant versions of genes that are non-functional. This is accomplished by altering the gene itself and subsequently bringing both copies of a non-functional gene together in one plant. This strategy works for most of the ~30,000 Arabidopsis genes, which are present in the genome at single copy. However, roughly 8,000 Arabidopsis genes exist as families of closely related copies that are much more difficult to coordinately inactivate by conventional methods. The goal of this CSHL 2010 project is to develop resources that allow the function of the genes that form these multi-gene families to be determined. To accomplish this, the project will take advantage of a conserved biological pathway that acts as a programmable engine for gene silencing. This pathway is known as RNA interference or RNAi. In response to double-stranded RNA, RNAi turns off the expression of any gene that shares sequence with the silencing trigger. Unlike conventional mutants, one copy of an artificial gene expressing double-stranded RNA is sufficient to silence all members of even a multi-gene family. Moreover, double-stranded RNA production can be regulated in a manner that restricts the inactivation of genes to specific tissues or times in development. RNAi triggers against each of the 8000 genes that are present as members of Arabidopsis multi-gene families will be produced. Additionally, RNAi triggers against genes that are essential early in development will be created, as the ability to turn off the activity of these genes in selected tissues or in a timed manner may reveal aspects of their function that cannot be illuminated by conventional mutations. Resources developed through this project will complement existing mutant collections, as conventional tools are not suited to determining the function of genes within multi-gene families and genes with early lethal phenotypes. Without such complementary approaches, the goal of understanding the function of all genes in Arabidopsis by 2010 will be unattainable. All resources will be made readily available to the scientific community as they are developed through a public repository, the Arabidopsis Biological Resource Center and through a commercial reagent distribution network, Open Biosystems. Access to information regarding the status of the resource, the overall project and how to obtain materials can be obtained from the project website at http://2010.cshl.edu. Information about the participating investigators and their research programs are also available. With respect to outreach and training, the project will leverage efforts at Cold Spring Harbor Laboratories through the Dolan DNA Learning Center to incorporate the plant resources generated into their curriculum.