Systematic Vertebrate Functional Genomics

 描述（由申请人提供）：脊椎动物基因组包含预测的20，000+个基因，许多基因的生物学作用未知。此外，这些分子中的大部分在不同的过程中具有不同的功能。脊椎动物基因组的功能注释是这个长期研究计划的首要目标。例如，即使在人类基因组工作完成十年后，正在进行积极研究的基因的多样性也没有发生实质性变化。我们开发了一系列基因断裂转座子（GBT）等位基因，用于使用杰出的非哺乳动物模式生物斑马鱼（斑马鱼）的脊椎动物基因组的表型注释。为什么强调蛋白质组？基因组学方法已经有力地表征了核基因组和转录组。然而，在动态背景下和体内的蛋白质组的完整复杂性的基因组评估仍然是未知的。斑马鱼提供了对整个脊椎动物蛋白质组进行全面分析的机会。我们将使用我们正在进行的700多个GBT收集和新的靶向TALEN/CRISPR敲除等位基因来评估脊椎动物表型组。在这次竞争性更新中，我们将专注于生物功能的注释，以识别开发中的新参与者以及一组临床相关的生物过程。我们将通过以下具体目标实现这一目标：具体目标一。我们将使用斑马鱼注释脊椎动物发育过程中的线粒体基因。具体目标二。我们将注释在胚胎和成人皮肤中表达和功能的新基因，并分析它们对器官稳态和再生潜力的贡献。具体目标三：我们将确定心脏突变体的注释基因修饰的成人心肌病。具体目标四。我们将鉴定消化器官突变，以表征各种潜在致病突变株系在脂质摄取、代谢、运输和储存方面的改变。我们选择了1）线粒体生物学，因为这个细胞器在正常生物学和疾病中的复杂作用，以及从核编码线粒体基因胚胎中的第一个GBT等位基因中意外发现的不同表型2）由于皮肤病对三分之一的美国人的强烈影响，珠被发育和功能3）心脏生物学和功能，因为心脏病是美国的主要死亡原因，4）脂质生物学和消化系统疾病生理学，因为它们的关键作用 心脏病和肥胖症。总之，该计划将为新的脊椎动物发育过程和对健康至关重要的生理系统的遗传基础提供新的见解，并且当改变时会导致疾病。

 DESCRIPTION (provided by applicant): The vertebrate genome contains a predicted 20,000+ genes, many of unknown biological role(s). In addition, a large fraction of these molecules have distinct functions in diverse processes. The functional annotation of the vertebrate genome is the overarching goal of this long-standing research program. For example, even a decade after the completion of the human genome effort, the diversity of genes undergoing active research has not substantially changed. We developed a collection of gene-break transposon (GBT) alleles for use in phenotypic annotation of the vertebrate genome using the preeminent non-mammalian model organism, the zebrafish (Danio rerio). Why emphasize the proteome? Genomic approaches have robustly characterized the nuclear genome and transcriptome. However, the genomic assessment of the full complexity of the proteome in a dynamic context and in vivo is still largely unknown. The zebrafish provides an opportunity to perform a comprehensive analysis of the entire vertebrate proteome. We will use our ongoing 700+ GBT collection and new, targeted TALEN/CRISPR knockout alleles for assessment of the vertebrate phenome. In this competitive renewal, we will focus on the annotation of biological function to identify new players in development as well as a set of clinically relevant biological processes. We will accomplish this goal through the following specific aims: Specific Aim I. We will annotate nuclearly encoded mitochondrial genes during vertebrate development using zebrafish. Specific Aim II. We will annotate novel genes with expression and function in the embryonic and adult skin and will analyze their contribution to organ homeostasis and regenerative potential. Specific Aim III: We will identify cardiac mutants for annotation of gene modifiers of adult cardiomyopathy. Specific Aim IV. We will identify digestive organ mutations to characterize a variety of potentially disease-causing mutant lines for alterations in lipid uptake, metabolism, transport and storage. We selected 1) Mitochondrial biology due to the complex role of this organelle in normal biology and disease and the unexpected finding of diverse phenotypes from the first GBT alleles in nuclearly encoded mitochondrial genes embryonic 2) Integument development and function due to the strong impact of skin disease on one in three Americans 3) Cardiac biology and function due to the critical role of heart disease as the leading cause of death in the US and 4) Lipid biology and digestive disease physiology due to their critical role(s) in development, heart disease and obesity. Together, this program will provide new insights into the genetic basis of new vertebrate developmental processes and physiological systems critical for health and that when altered contribute to disease.