Control of C. elegans Lineage by Heterochronic Genes

异时基因对线虫谱系的控制

• 批准号: 7081421
• 负责人: GARY B RUVKUN
• 金额: $ 53.68万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-05-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7081421
• 关键词: Caenorhabditis elegans; RNA binding protein; animal genetic material tag; biochemical evolution; bioinformatics; developmental genetics; fusion gene; gene deletion mutation; gene expression; genetic mapping; genetic regulatory element; genetic translation; genetically modified animals; green fluorescent proteins; helminth genetics; invertebrate embryology; messenger RNA; microRNAs; molecular cloning; neural transmission; phenotype; polymerase chain reaction; polysomes; protein structure function; regulatory gene; small interfering RNA

DESCRIPTION (provided by applicant): MicroRNAs have been discovered in a wide range of eukaryotes and at 21 to 24 nt are the smallest genes discovered in any organism. The first two miRNAs, lin-4 and let-7, emerged from the genetic analysis of the C. elegans developmental timing pathway. These miRNAs downregulate via complementary 3' UTR elements the translation of specific target mRNAs revealed by genetic analysis. The let-7 miRNA was the first shown to be conserved in animal phylogeny. Other tiny RNAs, the siRNAs, emerged from the biochemical analysis of RNA interference (RNAi). A common biochemical pathway processes both the siRNAs and miRNAs: the same Dicer RNAse and members of the PIWI class of proteins. Now hundreds of miRNAs have been discovered biochemically and by informatics, many of which are conserved across animal phylogeny. In this proposal we explore using RNAi the components of the pathways that process and present miRNAs and siRNAs to target mRNAs. We also explore the particular functions of specific miRNAs that are conserved in animal phylogeny we will fuse a number of conserved miRNA genes to GFP to reveal patterns of expression. We will observe the phenotypes of the deletion mutant strains for particular miRNA genes to explore their function. Candidate target mRNAs will be identified by bioinformatics and RNAi inactivated to test their action the pathway. We will also explore the function of miRNAs in the nervous system. We will use RNAi and genetics to discover genes that are necessary for miRNA repression of translation and siRNA induced mRNA degradation. We will test whether the proteins so identified are in complexes with miRNAs and target mRNAs. We will also genetically study the process by which RNAi is negatively regulated. We will explore whether miRNAs can act as signals between cells.

描述（由申请人提供）：在各种真核生物中发现了microRNA，在21至24 nt中是在任何生物体中发现的最小基因。前两个miRNA lin-4和let-7来自秀丽隐杆线虫发育时序途径的遗传分析。这些miRNA通过互补的3'UTR元素下调，遗传分析揭示了特定靶标mRNA的翻译。 Let-7 miRNA是在动物系统发育中首次显示的。其他细小的RNA，siRNA，来自RNA干扰的生化分析（RNAi）。一种常见的生化途径处理siRNA和miRNA：相同的丁香酶和Piwi类蛋白质的成员。现在，通过生化和信息学发现了数百个miRNA，其中许多是在动物系统发育中保守的。在此提案中，我们使用RNAi探索了对途径的组成部分，并呈现miRNA和siRNA以靶向mRNA。我们还探索了在动物系统发育中保守的特定miRNA的特定功能，我们将融合许多保守的miRNA基因与GFP，以揭示表达模式。我们将观察特定miRNA基因的缺失突变菌株的表型以探索其功能。候选目标mRNA将通过生物信息学识别，RNAi灭活以测试其动作途径。我们还将探索miRNA在神经系统中的功能。我们将使用RNAi和遗传学来发现对miRNA抑制翻译和siRNA诱导mRNA降解所必需的基因。我们将测试如此确定的蛋白质是否与miRNA和靶mRNA中的复合物中。我们还将基因研究RNAi受负调控的过程。我们将探索miRNA是否可以充当细胞之间的信号。