Small RNAs in skeletogenesis

骨骼发生中的小RNA

• 批准号: 7391638
• 负责人: TATSUYA KOBAYASHI
• 金额: $ 17.97万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2009-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7391638
• 关键词: Binding; Biochemical; Biological Process; Bone Development; Cartilage; Cell Proliferation; Cells; Chondrocytes; Chromatin; Class; Condition; Data; Databases; Development; Disease; Disruption; Endoribonucleases; Exons; Family; Functional RNA; Gene Expression; Gene Expression Regulation; Gene Targeting; Generations; Genes; Genetic Recombination; Goals; In Situ Hybridization; Informatics; Knowledge; Length; Mammalian Cell; Mammals; Methods; MicroRNAs; Mus; Mutant Strains Mice; Nucleotides; Organism; Osteogenesis; Pancreatic ribonuclease; Pattern; Physiological; Plants; Play; Polymerase Chain Reaction; Population; Regulation; Ribonuclease III; Ribonucleases; Role; Screening procedure; Skeletal Development; Skeletal system; Small Interfering RNA; Small RNA; Staging; System; Tissues; Transgenic Mice; Translating; Validation; Zebrafish; base; cell type; fetal; fly; gene cloning; gene therapy; human DICER1 protein; in vivo; mouse genome; novel therapeutics; postnatal; recombinase; research study; therapeutic target

DESCRIPTION (provided by applicant): Small RNAs (sRNAs) constitute a family of regulatory non-coding RNAs of up to 30 nucleotides in length. Two major classes of endogenous small RNAs, microRNAs (miRNAs) and endogenously occurring small interfering RNAs (siRNAs) negatively regulate gene expression by binding target mRNAs and by guiding chromatin silencing machineries, respectively. These sRNAs have been demonstrated to play critical roles in diverse biological processes in various organisms such as plants, worms, flies, and zebrafish. However, despite the abundant presence of miRNAs, knowledge about roles of sRNAs in mammals, let alone in mammalian skeletal tissues, is limited. The goals of this proposed project are first, to determine whether sRNAs play physiologically important roles in skeletal tissues in vivo, and second, to determine expression patterns and levels of sRNAs in skeletal tissues. In Aim I, we will delete the gene encoding Dicer, an RNase essential for generation of mature miRNA and siRNA, in mouse cartilage using the Cre-/oxP recombination system. Skeletal abnormalities of mice missing Dicer will be analyzed to define the effect of loss of miRNAs and siRNAs. This experiment should establish physiological roles of Dicer-dependent sRNAs during skeletal development. In Aim II, we will analyze expression patterns and levels of miRNAs, the predominant population of sRNAs in mammals. We will use miRNA array analysis for screening, quantitative PCR and in situ hybridization for validation and further characterization of each miRNA selected from the screening. Experiments in Aim II should provide a baseline database for miRNA expression in cartilage, which would help to form hypotheses about roles of specific miRNAs and their target genes in skeletal development. Understanding roles of sRNAs in skeletal tissues would not only reveal another level of regulation of gene expression, but also may provide novel therapeutic targets for genetic intervention in disease conditions.

描述（由申请人提供）：小RNA（sRNA）构成了长度最多30个核苷酸的调节性非编码RNA家族。内源性小 RNA 的两大类，即 microRNA (miRNA) 和内源性小干扰 RNA (siRNA)，分别通过结合靶 mRNA 和引导染色质沉默机制来负向调节基因表达。这些 sRNA 已被证明在植物、蠕虫、苍蝇和斑马鱼等多种生物体的多种生物过程中发挥着关键作用。然而，尽管 miRNA 大量存在，但关于 sRNA 在哺乳动物中的作用的知识仍然有限，更不用说在哺乳动物骨骼组织中了。该项目的目标首先是确定 sRNA 是否在体内骨骼组织中发挥重要的生理作用，其次是确定 sRNA 在骨骼组织中的表达模式和水平。在目标 I 中，我们将使用 Cre-/oxP 重组系统删除小鼠软骨中编码 Dicer 的基因，Dicer 是生成成熟 miRNA 和 siRNA 所必需的 RNase。将分析缺少 Dicer 的小鼠骨骼异常，以确定 miRNA 和 siRNA 丢失的影响。该实验应确定 Dicer 依赖性 sRNA 在骨骼发育过程中的生理作用。在 Aim II 中，我们将分析 miRNA（哺乳动物中 sRNA 的主要群体）的表达模式和水平。我们将使用 miRNA 阵列分析进行筛选，使用定量 PCR 和原位杂交对筛选中选择的每个 miRNA 进行验证和进一步表征。 Aim II 中的实验应该提供软骨中 miRNA 表达的基线数据库，这将有助于形成关于特定 miRNA 及其靶基因在骨骼发育中的作用的假设。了解 sRNA 在骨骼组织中的作用不仅可以揭示基因表达的另一个调控水平，还可以为疾病的遗传干预提供新的治疗靶点。

项目成果

MicroRNAs in cartilage development, homeostasis, and disease.

• DOI: 10.1007/s11914-014-0229-9
• 发表时间: 2014-12
• 期刊: CURRENT OSTEOPOROSIS REPORTS
• 影响因子: 4.3
• 作者: Mirzamohammadi, Fatemeh;Papaioannou, Garyfallia;Kobayashi, Tatsuya
• 通讯作者: Kobayashi, Tatsuya

MicroRNA-140 Provides Robustness to the Regulation of Hypertrophic Chondrocyte Differentiation by the PTHrP-HDAC4 Pathway.

• DOI: 10.1002/jbmr.2438
• 发表时间: 2015-06
• 期刊: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research
• 作者: Papaioannou G;Mirzamohammadi F;Lisse TS;Nishimori S;Wein MN;Kobayashi T
• 通讯作者: Kobayashi T