piRNA biogenesis and function in germline development

piRNA 生物发生和种系发育中的功能

基本信息

批准号：
8212182
负责人：
WILLIAM Edward THEURKAUF
金额：
$ 58.06万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-12-01 至 2015-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8212182
关键词：
Address Aubergine Binding Biochemical Biogenesis Biological Process ChIP-seq Chromosome Deletion Chromosome Segregation Chromosomes Cleaved cell Complex Computational algorithm Computer Analysis Cytoplasm DNA-Directed RNA Polymerase Data Development Drosophila genus Elements Embryo Embryonic Development Epigenetic Process Fishes Genetic Genetic Transcription Genetic Translation Genome Genomics Goals Hereditary Disease Homologous Gene Human Genome In Vitro Link Location Maintenance Malignant Neoplasms Measures Meiosis Mitotic Mitotic Chromosome Mobile Genetic Elements Molecular Molecular Genetics Mus Mutation Nucleotides Oogenesis Pathway interactions Pattern Production Property Protein Binding Proteins RNA RNA Interference RNA Sequences Reporter Retrotransposon Role Site Small RNA System Testing Transcript Transgenic Organisms chromatin immunoprecipitation fly genome-wide histone modification human DICER1 protein improved in vivo insight mutant novel public health relevance research study telomere

项目摘要

DESCRIPTION (provided by applicant): The PIWI interaction RNA (piRNA) pathway is required for transposon silencing during germline development in flies, fish and mouse. The 24 to 29 nucleotide long piRNAs are produced by a Dicer-independent mechanism and associate specifically with PIWI class Argonaute proteins. These piRNA-PIWI complexes can cleave target RNAs in vitro. However, the in vivo mechanism of transposon silencing and the genomic origins of piRNAs are not understood. In addition, the full spectrum of biological functions for the piRNA pathway remains to be explored. These broad questions will be addressed using a combination of genetic, genomic, cytological and molecular approaches in the experimentally tractable Drosophila system. The broad aims of the proposal are to define the structural and biochemical properties of piRNA encoding clusters, determine the mechanism of transposon silencing by piRNAs, and to characterize recently identified functions for this pathway in meiotic and mitotic chromosome segregation. Studies under aim 1 will use computational analysis of deep sequencing data and chromatin immunoprecipitation to define properties of piRNA encoding clusters. Aim 2 will use transgenic reporters to explore the mechanism of piRNA silencing. Studies under Aim 3 will use genetic, molecular and cytological approaches to define the role of piRNA in telomere protection and chromosome segregation. PUBLIC HEALTH RELEVANCE: Transposon are mobile genetic elements that make up almost 50% of the human genome. Mobilization of these elements can cause genetic diseases, including cancer. piRNAs have a conserved function in maintaining genome integrity and suppressing transposon mobilization. The long term goal of the proposed studies is to understand how piRNA controls the transposon activity.

描述（由申请人提供）：果蝇、鱼和小鼠种系发育过程中转座子沉默需要 PIWI 相互作用 RNA (piRNA) 途径。 24 至 29 个核苷酸长的 piRNA 由 Dicer 独立机制产生，并与 PIWI 类 Argonaute 蛋白特异性关联。这些 piRNA-PIWI 复合物可以在体外切割靶 RNA。然而，转座子沉默的体内机制和 piRNA 的基因组起源尚不清楚。此外，piRNA 途径的全部生物学功能仍有待探索。这些广泛的问题将在实验上可处理的果蝇系统中结合使用遗传、基因组、细胞学和分子方法来解决。该提案的主要目标是定义 piRNA 编码簇的结构和生化特性，确定 piRNA 转座子沉默的机制，并表征最近确定的该途径在减数分裂和有丝分裂染色体分离中的功能。目标 1 下的研究将使用深度测序数据和染色质免疫沉淀的计算分析来定义 piRNA 编码簇的特性。目标2将使用转基因报告基因来探索piRNA沉默的机制。目标 3 下的研究将使用遗传、分子和细胞学方法来定义 piRNA 在端粒保护和染色体分离中的作用。公共卫生相关性：转座子是可移动的遗传元件，几乎占人类基因组的 50%。这些元素的动员可能导致遗传疾病，包括癌症。 piRNA 在维持基因组完整性和抑制转座子动员方面具有保守功能。拟议研究的长期目标是了解 piRNA 如何控制转座子活性。