SysCODE: PMAGE Technology Development (10 of 10)

SysCODE：PMAGE 技术开发（10 条，共 10 条）

基本信息

批准号：
7883587
负责人：
JONATHAN G SEIDMAN
金额：
$ 25.43万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-30 至 2012-06-30
项目状态：
已结题

项目摘要

used to define pathways in mammalian organ development. Because the central focus of the SysCODE Consortium is to define such pathways in the developing tooth germ, pancreatic islet and heart valve, Projects 2, 3 and 4 of this U54 Consortium will use RNA profiling to define RNA levels during development. We have recently developed a novel RNA profiling procedure, PMAGE, that circumvents potential issues encountered with conventional microarray platforms in the analysis of low abundance transcripts. The PMAGE method combines the frequency distribution technique pioneered by SAGE (serial analysis of gene expression) with the high throughput Polony DNA sequencing technique to produce histograms, which accurately profile the RNAs in tissues or cells. That is, a single 14 base pair tag is produced from a single RNA molecule, the tags are stored in PMAGE libraries, and about 2 million tags are sequenced per library. The RNA profiles obtained by this method contain about 30-fold more tags than the majority of SAGE libraries, and hence are able to measure the level of RNAs expressed at less than 0.3 RNA copies per cell. This enhanced sensitivity thus encompasses potential low abundance RNAs, such as those encoding transcription factors and other regulatory molecules that may be important to a molecular understanding of organogenesis. The ability to comprehensively define levels of transcription factors should therefore provide novel insights into organ development. We propose to create, under the aegis of this P30 Core grant, a PMAGE Technology Development Core facility that will enable Consortium investigators to determine expression profiles from total RNA. We expect to produce about 50 PMAGE RNA profiles in the first year and 250 RNA profiles during the grant period. These RNA profiles will provide unique insights into the mechanisms by which the tooth germ, pancreatic islet and heart valve form. In particular we will define the complete catalogue of transcription factors that are required for the development of these tissues. PERFORMANCE SITE(S) (organization, city, state) Dept of Genetics, Harvard Medical School, Boston, MA 02115 PHS 398 (Rev. 09/04) Page 2 Form Page 2

用于定义哺乳动物器官发育的途径。因为 SysCODE 的核心焦点联盟将定义牙胚、胰岛和心脏瓣膜发育过程中的此类通路， U54 联盟的项目 2、3 和 4 将使用 RNA 分析来定义开发过程中的 RNA 水平。我们最近开发了一种新颖的 RNA 分析程序 PMAGE，可以避免潜在的问题在分析低丰度转录本时遇到了传统的微阵列平台。这 PMAGE方法结合了SAGE首创的频率分布技术（基因序列分析）表达）与高通量Polony DNA测序技术产生直方图，准确地分析组织或细胞中的 RNA。也就是说，单个 14 碱基对标签是由单个 RNA分子的标签存储在PMAGE文库中，每个文库对大约200万个标签进行测序。通过这种方法获得的 RNA 图谱包含的标签比大多数 SAGE 多约 30 倍文库，因此能够测量每个细胞表达少于 0.3 个 RNA 拷贝的 RNA 水平。因此，这种增强的灵敏度涵盖了潜在的低丰度 RNA，例如编码的 RNA 转录因子和其他调节分子可能对分子理解很重要器官发生。因此，全面定义转录因子水平的能力应该提供对器官发育的新见解。我们建议在 P30 核心拨款的支持下创建一个 PMAGE 技术开发核心设施将使联盟研究人员能够确定总 RNA 的表达谱。我们预计第一年会产生约 50 个 PMAGE RNA 图谱以及资助期间的 250 个 RNA 概况。这些 RNA 谱将为我们提供独特的见解牙胚、胰岛和心脏瓣膜的形成机制。特别是我们将定义这些组织发育所需的转录因子的完整目录。绩效站点（组织、城市、州）哈佛医学院遗传学系，波士顿，MA 02115 PHS 398（修订版 09/04）第 2 页表格第 2 页