Systematic Vertebrate Functional Genomics

描述(申请人提供)：脊椎动物基因组预测包含100,000个基因，大部分功能未知。的最新发展 基于吗啡的斑马鱼基因敲除技术为 在模式脊椎动物中，基于序列的全基因组功能分配。 我们建议探索控制基础的分子机制。 脊椎动物胚胎的图案化和器官形成过程，生物学 细胞间通讯在发育过程中至关重要的问题。我们 我将把这项研究的重点放在系统地将生物功能分配给 一组100-200个假定的分泌性蛋白质以识别这些关键角色 细胞间胚胎信号级联。 我们将通过以下具体目标实现这一目标： 目的1.建立分泌型吗啉蛋白核心数据库。基因 编码预测的分泌蛋白将通过分析 斑马鱼基因组和EST数据库。针对这些基因的Morpholinos将 产生并检查在下列生物过程中的功能： 目的II.分离早期胚胎和神经所需的分泌蛋白 图案化。目的III.分离心血管所需的分泌蛋白 和感觉器官的形成。目的IV.所需的分泌蛋白的分离 用于消化器官的形成和功能。 脊椎动物构型和器官所需分子的鉴定 形成对基因的理解具有重要的意义 这些过程中的不足之处。此外，这些信号蛋白，至关重要 对于体内发育来说，将是建立条件的关键 体外器官形成。斑马鱼提供了第一个全面的分析 使用整个脊椎动物基因组作为模板。

DESCRIPTION (provided by applicant): The vertebrate genome contains a predicted 100,000 genes, most of unknown function. The recent development of morpholino-based gene knockdown technology in zebrafish has opened the door to the genome-wide assignment of function based on sequence in a model vertebrate. We propose to explore the molecular mechanisms that control fundamental vertebrate embryonic processes of patterning and organ formation, biological problems in which cell-cell communication is critical during development. We will focus this study on the systematic assignment of biological function to a set of 100-200 putative secreted proteins to identify key players in these intercellular embryonic signaling cascades. We will accomplish this goal through the following specific aims: Aim I. Development of a secreted protein morpholino core database. Genes encoding predicted secreted proteins will be selected through analysis of the zebrafish genomic and EST database. Morpholinos targeted to these genes will be generated and examined for function in the following biological processes: Aim II. Isolation of secreted proteins required for early embryonic and neural patterning. Aim III. Isolation of secreted proteins required for cardiovascular and sensory organ formation. Aim IV. Isolation of secreted proteins required for digestive organ formation and function. The identification of molecules required for vertebrate patterning and organ formation has critical implications for the understanding of genetic deficiencies in these processes. In addition, these signaling proteins, crucial for development in vivo, will be crucial to the establishment of conditions for in vitro organ formation. The zebrafish offers the first comprehensive analysis of these processes using as template an entire vertebrate genome.