Regulatory Roles of miRNAs during Early Arabidopsis Embryogenesis

miRNA 在早期拟南芥胚胎发生过程中的调控作用

• 批准号: 7545748
• 负责人: Michael D Nodine
• 金额: $ 4.68万
• 依托单位: WHITEHEAD INSTITUTE FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7545748
• 关键词: Anabolism; Animals; Arabidopsis; Biogenesis; Cell Differentiation process; Cells; Defect; Development; Developmental Process; Disease; Embryo; Embryonic Development; Event; Exhibits; Gene Expression; Goals; In Situ Hybridization; Laser Scanning Confocal Microscopy; Localized; Mediating; MicroRNAs; Microscopic; Molecular Genetics; Morphogenesis; Mouse-ear Cress; Nucleotides; Oligonucleotide Probes; Pattern; Pattern Formation; Phenocopy; Plants; RNA; Regulation; Research; Research Design; Role; Small RNA; Staging; Standards of Weights and Measures; System; Techniques; Technology; Testing; Transcript; Transgenic Organisms; Untranslated RNA; base; blastomere structure; cell type; human disease; in vivo; insight; laser capture microdissection; locked nucleic acid; mutant; programs; research study; sample fixation; sensor

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) are -21 nucleotide noncoding RNAs that posttranscriptionally regulate gene expression in plants and animals. Although miRNA-mediated regulation is clearly essential for proper morphogenesis and differentiation in plants and animals, relatively little is known about the mechanistic basis of miRNA functions during development. Arabidopsis thaliana embryos are an ideal system for characterizing the developmental roles of miRNAs for several reasons, including: 1) miRNAs have important functions during Arabidopsis embryogenesis, 2) the vast majority of Arabidopsis miRNA targets have been confidently predicted, 3) molecular genetic approaches can be efficiently used to characterize miRNA/target interactions in vivo, and 4) there are several advantages to studying pattern formation in Arabidopsis embryos. The proposed research plan therefore utilizes Arabidopsis embryos to characterize the roles of miRNAs in fundamental cell fate decisions with the long-term goal of understanding the mechanistic basis of miRNAs during development. The specific aims of the proposed research are designed to assess the regulatory roles of miRNAs during early embryonic differentiation events and, more generally, to test whether miRNAs initiate, maintain and/or fine-tune cell differentiation programs. By testing these hypotheses, insights into the functions of miRNAs in basic developmental processes will be generated. Cell-specific markers will first be examined in embryos with defects in miRNA biogenesis to identify the earliest stages and differentiation events that miRNAs are involved in. The corresponding miRNAs required for these cell-fate decisions will be identified using laser capture microdissection and highthroughput sequencing technologies. The localization patterns and functional domains of these early embryonic miRNAs will be characterized using RNA in situ hybridizations and miRNA sensor constructs. Candidate targets of early embryonic miRNAs will then be identified by determining which target transcripts are mis-regulated in miRNA biosynthesis mutants. The in vivo functions of miRNA/target interactions will be assessed using molecular genetic approaches. Characterizing the regulatory roles of miRNAs during cellular differentiation will contribute to a better understanding of miRNA functions. Since miRNA mis-regulation has been implicated in human disease, the proposed research may therefore yield insight into the mechanistic basis of disease-afflicted cell types.

描述（由申请人提供）：microRNA（miRNA）是-21核苷酸非编码RNA，在转录后调节植物和动物的基因表达。尽管miRNA介导的调节对于植物和动物的适当形态发生和分化显然至关重要，但对发育过程中miRNA功能的机械基础知之甚少。拟南芥胚胎是出于几个原因表征miRNA的发育作用的理想系统，包括：1）miRNA在拟南芥胚胎发生期间具有重要功能，2）绝大多数拟南芥miRNA靶标已有信心地预测，3）有多种图案可以有效地进行mirna/tarties contrumitions and Mirna互动，并在4个构图中，4. 4拟南芥胚胎中的形成。因此，拟议的研究计划利用拟南芥胚胎来表征miRNA在基本细胞命运决策中的作用，其长期目标是理解发育过程中miRNA的机理基础。拟议研究的具体目的旨在评估早期胚胎分化事件中miRNA的调节作用，更普遍地测试miRNA是否启动，维护和/或微调细胞分化程序。通过检验这些假设，将产生对miRNA在基本发展过程中的功能的见解。首先将在胚胎中检查细胞特异性标记，并在miRNA生物发生中存在缺陷，以识别miRNA所涉及的最早阶段和分化事件。这些细胞污染决策所需的相应miRNA将使用激光捕获的微解析和高促进测序技术来识别。这些早期胚胎miRNA的定位模式和功能结构域将使用原位杂交和miRNA传感器构建体进行表征。然后，将通过确定哪些目标转录物在miRNA生物合成突变体中错误调节的候选靶标的候选靶标。 miRNA/靶标相互作用的体内功能将使用分子遗传方法评估。表征miRNA在细胞分化过程中的调节作用将有助于更好地理解miRNA功能。由于miRNA的错误调节已与人类疾病有关，因此拟议的研究可能会深入了解疾病伴性细胞类型的机械基础。