SysCODE: PMAGE Technology Development (10 of 10)

SysCODE：PMAGE 技术开发（10 条，共 10 条）

• 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
• 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
• 来源: https://reporter.nih.gov/project-details/7883587
• 关键词: Adult; Base Pairing; Binding; Biomedical Research; Boston; Cancer Center Support Grant; Cardiac; Cataloging; Catalogs; Cells; Cities; Complementary DNA; Core Facility; DNA Sequence; Development; Engineering; Expression Library; Frequencies; Gene Expression; Gene Expression Profiling; Genes; Genetic; Grant; Heart; Heart Valves; In Vitro; Islets of Langerhans; Left ventricular structure; Libraries; Ligation; Manuscripts; Measures; Methods; Microscope; Molecular; Molecular Profiling; Mus; Nucleotides; Organ; Organogenesis; Pathway interactions; Pattern; Procedures; RNA; Research Personnel; Sampling; Slide; Techniques; Technology; Tissue Sample; Tissues; Tooth Germ; Transcript; base; cost efficient; improved; insight; medical schools; novel; performance site; ranpirnase; research study; serial analysis of gene expression; technology development; tool; transcription factor

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中的途径。因为Syscode的主要重点 财团将在发育中的牙齿胚芽，胰岛和心脏瓣膜中定义此类途径， 该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型的RNA分析程序Pmage，该程序绕过潜在问题 在低丰度转录本的分析中，与常规的微阵列平台相遇。这 PMAGE方法结合了通过SAGE开创的频率分布技术（基因的序列分析 用高吞吐量的polpol dna测序技术表达），以产生直方图 准确介绍组织或细胞中的RNA。也就是说，一个14个基对标签是由单个产生的 RNA分子，标签存储在PMAGE库中，每个库进行了约200万个标签。 通过这种方法获得的RNA曲线包含的标签比大多数Sage多30倍 库，因此能够测量每个细胞以小于0.3 RNA拷贝表达的RNA水平。 因此，这种增强的灵敏度涵盖了潜在的低丰度RNA，例如编码的敏感性 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提供 对器官发育的新颖见解。我们建议在此P30核心赠款的宙斯盾下创建一个 PMAGE Technology Development核心设施将使财团调查员确定 总RNA的表达谱。我们预计第一年将产生约50个PMAGE RNA概况 和250个RNA概况在赠款期间。这些RNA配置文件将为您提供独特的见解 牙齿胚芽，胰岛和心脏瓣膜形式的机制。特别是我们将定义 这些组织开发所需的转录因子的完整目录。 绩效网站（组织，城市，州） 马萨诸塞州波士顿哈佛医学院遗传学系02115 PHS 398（修订版09/04）第2页表格页2