RNA Biology in Oxytricha: a Complex Unicellular Model

尖毛虫中的 RNA 生物学：复杂的单细胞模型

• 批准号: 8888050
• 负责人: LAURA F LANDWEBER
• 金额: $ 30.4万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8888050
• 关键词: Animal Model; Attention; Biogenesis; Bioinformatics; Biological Models; Biology; Chromosomal Rearrangement; Chromosomes; Complex; DNA; DNA Sequence; Data; Development; Developmental Biology; Disease; Eukaryota; Event; Frequencies; Funding; Gene Expression; Gene Rearrangement; Generations; Genes; Genetic; Genetic Recombination; Genetic Transcription; Genome; Genome Stability; Genomic Instability; Goals; Human; Inherited; Knowledge; Lead; Light; Maintenance; Malignant Neoplasms; Mediating; Messenger RNA; Modeling; Molecular; Nuclear; Nucleic Acid Regulatory Sequences; Organism; Oxytricha; Pathway interactions; Process; Production; Property; RNA; RNA Splicing; Regulation; Research; Small RNA; System; Testing; Tumor Suppressor Genes; United States National Institutes of Health; Untranslated RNA; Ursidae Family; cancer genome; chimeric gene; chromothripsis; experimental analysis; genome integrity; genome sequencing; genome-wide; human disease; insight; knockout gene; microbial; multidisciplinary; next generation; novel; piRNA; programs; public health relevance; research study; tool; tumor

 DESCRIPTION (provided by applicant): RNA molecules display a novel mode of action in pond-dwelling ciliated protists, where both long and small noncoding RNAs regulate an intricate process of genome remodeling. Maternally-inherited piRNA molecules in Oxytricha mark which 225,000 segments of the genome to retain, and long, template RNAs program the order of those DNA segments during massive genome rearrangements. The proposed goal of this research is to understand the biogenesis and regulation of both these long and small noncoding RNAs, as well as the regulation of other steps in pathways of RNA production in the model organism Oxytricha. Expanding our suite of tools for studying and manipulating this emerging genetic system is also a high priority, and one aim will capitalize on our knowledge of piRNA biology in these organisms to develop a powerful tool for gene knockout. This will, in turn, help us better understand other aspects of the genetics and developmental biology of genome rearrangements, as well as its implications for the maintenance and regulation of genome integrity, more generally. The proposed experiments will examine the properties of both the molecules that participate in these complex rearrangement events and the machinery underlying them. The most immediate insight to be gained from the proposed research is an understanding of the range of activities accessible to both long and small noncoding RNA molecules, and the piRNA pathway in Oxytricha, in particular, with attention to both differences and similarities between this system and other eukaryotic organisms. Specific aims include: 1. Studying piRNAs and their biogenesis in Oxytricha, 2. Develop piRNA-mediated gene knockout as the first gene knockout tool in Oxytricha, 3. Understand long, noncoding template RNA production and their interacting partners, 4. Identify and functionally test motifs that regulate gene expression from somatic chromosomes.

 描述（由申请人提供）： RNA分子在池塘栖息的纤毛原生生物中表现出一种新的作用模式，其中长非编码RNA和小非编码RNA调节基因组重塑的复杂过程。尖毛虫中母系遗传的 piRNA 分子标记了要保留的 225,000 个基因组片段，而长模板 RNA 在大规模基因组重排过程中对这些 DNA 片段的顺序进行编程。这项研究的拟议目标是了解这些长非编码 RNA 和小非编码 RNA 的生物发生和调控，以及模式生物尖毛虫中 RNA 生产途径中其他步骤的调控。扩展我们用于研究和操纵这一新兴遗传系统的工具套件也是一个高度优先事项，其中一个目标是利用我们在这些生物体中的 piRNA 生物学知识来开发一种强大的基因敲除工具。反过来，这将帮助我们更好地了解基因组重排的遗传学和发育生物学的其他方面，以及它对基因组完整性维护和调节的更广泛的影响。 拟议的实验将检查参与这些复杂重排事件的分子的特性及其背后的机制。从拟议的研究中获得的最直接的见解是了解长非编码 RNA 分子和小非编码 RNA 分子的活动范围，特别是尖毛虫中的 piRNA 途径，特别是注意该系统与其他真核生物之间的差异和相似之处。具体目标包括：1. 研究 piRNA 及其在尖毛虫中的生物发生，2. 开发 piRNA 介导的基因敲除作为尖毛虫中第一个基因敲除工具，3. 了解长非编码模板 RNA 的产生及其相互作用的伙伴，4. 识别和功能测试调节体细胞染色体基因表达的基序。