The Function of piRNAs and the piRNA Complex

piRNA 和 piRNA 复合物的功能

• 批准号: 7893419
• 负责人: NELSON C LAU
• 金额: $ 23.38万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-05 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7893419
• 关键词: Address; Animal Model; Animals; Architecture; Biochemical; Biogenesis; Biological Assay; Cell Line; Cell Nucleus; Cells; Cellular biology; Child health care; Chromatin; Chromatin Structure; Complex; DNA Transposable Elements; Development; Ectopic Expression; Elements; Family; Fertility; Fluorescent in Situ Hybridization; Foundations; Funding; Gene Expression Regulation; Gene Silencing; Genes; Genetic; Genetic Transcription; Genome; Genomics; Germ Cells; Goals; Health; Human; Human Development; Immunohistochemistry; Investigation; Knockout Mice; Link; Mammals; Methodology; Methods; MicroRNAs; Molecular; Mus; Oocytes; Organism; Pathway interactions; Process; Proteins; RNA Interference; RNA Interference Pathway; Rana; Rattus; Reagent; Regulation; Regulator Genes; Research; Research Project Grants; Ribonucleoproteins; Role; Small RNA; Stem cells; System; Techniques; Testis; Transcriptional Regulation; Transplantation; Untranslated RNA; Xenopus; career; chromatin immunoprecipitation; egg; fly; genome-wide; histone modification; insight; male; novel; programs; promoter; protein complex; tool

Small noncoding RNAs are central components ofthe gene regulatory circuit in organisms. For proper health and development, organisms employ protein complexes containing small endogenous RNAs to control thousands of other genes through silencing mechanism. Genetics in model organisms implicate a link between small RNAs, Argonaute proteins and processes that regulate chromatin structure, suggesting that RNA interference ^RNAi) pathways can impact large domains of the genome. Our search for an analogous link in mammals directed us to purify from rat testes the piRNA Complex (piRC), which consists at least of mammalian Piwi and the Piwi-interacting RNAs. Fiwi-interacting RNAs (piRNAs) are a diverse class of small RNAs that are longer than microRNAs and derive from intergenic clusters of between 1-lOOkb long. Piwi, a subclade of the Argonaute family, is essential for fertility in flies and male mice, and has been genetically implicated in silencing of transposable elements. Despite these features, a rigorous understanding of piRC's molecular function is still lacking. This proposal seeks to dissect the molecular function ofthe piRNAs and piRC through these specific aims: 1) Functional analysis ofthe piRNA Complex; 2) Regulation of piRNA cluster transcription; 3) Understanding the Genesis of piRNAs. We will apply our current biochemical techniques to probe piRC's functional activity, and develop genetic and cell biology methodologies to understand the role of piRNA clusters. Piwi proteins have been shown to be essential for proper germ cell development in animals, including mammals. However, the mechanistic understanding and gene regulatory potential ofthis ribonucleoprotein is still uncharted. This research project aims to develop systems that will drive progress in a subject relevant to child health and human development.

小非编码 RNA 是生物体基因调控回路的核心组成部分。为了正常健康和发育，生物体利用含有小内源 RNA 的蛋白质复合物通过沉默机制控制数千个其他基因。模式生物中的遗传学暗示了小RNA、Argonaute蛋白和调节染色质结构的过程之间的联系，这表明RNA干扰（^RNAi）途径可以影响基因组的大范围。我们在哺乳动物中寻找类似的联系，指导我们从大鼠睾丸中纯化 piRNA 复合物 (piRC)，它至少由哺乳动物 Piwi 和 Piwi 相互作用的 RNA 组成。 Fiwi相互作用RNA(piRNA)是多种小RNA，其比microRNA更长并且源自长度在1-100kb之间的基因间簇。 Piwi 是 Argonaute 家族的一个分支，对于果蝇和雄性小鼠的生育能力至关重要，并且与转座因子的沉默有关。尽管有这些特征，但仍然缺乏对 piRC 分子功能的严格理解。 该提案旨在通过以下具体目标剖析 piRNA 和 piRC 的分子功能：1）piRNA 复合物的功能分析； 2）piRNA簇转录的调控； 3) 了解 piRNA 的起源。我们将应用现有的生化技术来探测 piRC 的功能活性，并开发遗传和细胞生物学方法来了解 piRNA 簇的作用。 Piwi 蛋白已被证明对于动物（包括哺乳动物）生殖细胞的正常发育至关重要。然而，这种核糖核蛋白的机制理解和基因调控潜力仍然未知。该研究项目旨在开发能够推动儿童健康和人类发展相关主题取得进展的系统。