Arabidopsis 2010: Next Generation Cellular Resolution Profiling of the Transcriptome and Epigenome

拟南芥 2010：转录组和表观基因组的下一代细胞分辨率分析

• 批准号: 0929349
• 负责人: Elliot Meyerowitz
• 金额: $ 64万
• 依托单位: California Institute of Technology
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-15 至 2011-11-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0929349&HistoricalAwards=false
• 关键词: Arabidopsis; 2010; Next; Generation; Cellular

The purpose of the project is to develop and apply methods to sequence translating messenger RNA from specific cell types. An important deficit in current experiments that obtain the list of active genes in plant tissue is that the tissue used comes from a mixture of different cell types, for example, it uses whole flowers or whole seedlings, each of which is made up of many different types of cells. The resulting data on active genes is useful, but is information on the activity of genes from many cells that have different collections of active genes. As cell-type-specific data are necessary to infer gene regulatory networks, current types of data are inadequate to understand a complex organ like a leaf, shoot, or flower. The research will use developing flowers as a system, will adapt existing methods and develop new methods to isolate cell-type specific RNA, and to perform detailed sequencing of the RNA from individual cell types of the developing flower. The result will be a set of tested methods that allow researchers to isolate messenger RNA from single cell types found in complex tissues, and therefore to record patterns and changing patterns of gene activity from each of the single cell types found in developing organs.Broader ImpactsThe broader impacts of the project are in three areas. First, the completion of the proposed research will result in new technologies for the entire plant science community to study RNA and gene activity at cellular resolution in virtually any cell type, which is expected to be crucial for the understanding of various questions from development to energy capture and response to the environment. All the methods and lines generated through this project, which will enable cell-type specific analysis in other laboratories, including those of young and starting researchers who would have to spend considerable time to make such lines, will be made freely available to the research community. Second, by being partly established within the environment of a much larger group of undergraduate students, graduate students, postdoctoral fellows and technical staff that are engaged in different research projects on Arabidopsis and animal development (the Meyerowitz laboratory and the personnel of the Caltech Jacobs Center for Genetics and Genomics), the project will provide familiarity with a variety of genomic technologies and approaches to a substantial pool of future independent investigators. This pool will include undergraduate summer researchers and students from underrepresented minorities recruited through established programs at Caltech. Finally, public outreach and training activities will include provision of the materials generated (especially images) for the "Grounding in Botany" program, a collaboration between the Meyerowitz lab and the Huntington Botanical Gardens, in which laboratory exercises using plants are developed by teachers for high school classrooms; and for a program in which graduate students and postdoctoral scholars from Caltech spend up to six weeks at the Exploratorium in San Francisco, teaching high school and elementary science teachers in a summer teacher training program.

该项目的目的是开发和应用方法来对转换特定细胞类型的Messenger RNA的顺序进行测序。在当前实验中获得植物组织中活性基因列表的一个重要缺陷是，所使用的组织来自不同细胞类型的混合物，例如，它使用整个花朵或整个幼苗，每种幼苗由许多不同的幼苗组成细胞类型。在活性基因上产生的数据很有用，但是有关许多具有不同活性基因集合的基因活性的信息。 由于细胞类型的特异性数据对于推断基因调节网络是必需的，因此当前的数据类型不足以理解诸如叶子，芽或花朵之类的复杂器官。 该研究将使用开发花作为系统，将适应现有方法，并开发出新的方法来分离细胞类型的特异性RNA，并从发育中的花朵的单个细胞类型中对RNA进行详细的测序。 结果将是一组经过测试的方法，使研究人员能够从复杂组织中发现的单细胞类型中分离信使RNA，因此可以从开发器官中发现的每种单细胞类型记录基因活性的模式和变化的模式。该项目的更广泛影响是三个领域。首先，拟议的研究的完成将为整个植物科学界提供新的技术，以研究几乎任何细胞类型的细胞分辨率下的RNA和基因活性，这对于了解从发展到能量的各种问题至关重要捕获和对环境的反应。 通过该项目产生的所有方法和线条，将在其他实验室中进行细胞类型的特定分析，包括年轻的研究人员和初学者的研究人员，他们将不得不花费大量时间来制作此类行，并将其自由地提供给研究社区。其次，通过部分建立在更大的本科生，研究生，博士后研究员和技术人员的环境中，这些人员从事拟南芥和动物发展的不同研究项目（Meyerowitz实验室和Caltech雅各布斯中心的人员对于遗传学和基因组学），该项目将熟悉多种基因组技术和方法，以实现大量未来独立研究者。 该游泳池将包括通过加州理工学院既定计划招募的少数民族的本科生研究人员和来自代表性不足的少数民族的学生。 最后，公众推广和培训活动将包括提供“植物学基础”计划的生成的材料（尤其是图像），Meyerowitz Lab与亨廷顿植物园之间的合作高中教室；对于一个计划，该计划在夏季教师培训计划中，在旧金山的Exploratorium上最多六个星期的研究生和博士后学者在旧金山的Exploratorium呆了六个星期。