Post-transcriptional regulation in mammalian germ cell development

哺乳动物生殖细胞发育的转录后调控

• 批准号: 10090605
• 负责人: Donny D Licatalosi
• 金额: $ 32.04万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10090605
• 关键词: Address; Affect; Alternative Splicing; Animals; Binding; Binding Sites; Biochemical; Bioinformatics; Cell Differentiation process; Cell Nucleus; Cell Proliferation; Cells; Characteristics; Code; Cytoplasm; Data; Data Set; Defect; Development; Event; Extensive Stage; Fertility; Fluorescence; Fluorescence-Activated Cell Sorting; Gene Expression; Genes; Genetic; Genetic Transcription; Genetic Translation; Germ Cells; Goals; Growth; Heterogeneity; Human; Human Development; Incidence; Knockout Mice; Label; Laboratories; Length; Life; Maintenance; Mammalian Cell; Mammals; Maps; Measurement; Messenger RNA; Methods; Molecular; Mus; Mutation; Neonatal; Neurocognitive; Output; Paternal Age; Pathway interactions; Poly A; Polyadenylation; Positioning Attribute; Post-Transcriptional Regulation; Process; Production; Property; Protein Isoforms; Proteins; RNA; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Reproductive Biology; Research; Ribosomes; Role; Specific qualifier value; Spermatocytes; Stem Cell Development; Tail; Testing; Testis; TimeLine; Transgenic Mice; Translating; Untranslated RNA; Work; base; biochemical tools; bioinformatics tool; cancer cell; deep sequencing; germline stem cells; human disease; improved; innovation; insight; mRNA Expression; male; mouse model; offspring; postnatal; programs; recruit; sperm cell; tool; transcriptome

PROJECT SUMMARY The long-term goal of the proposed work is to gain a better understanding of how gene expression is regulated during mammalian cell development. This proposal focuses on the study of post-transcriptional control of gene expression, whereby mRNAs made from a single gene can be modified in different ways to change gene output. In the nucleus, this includes altering the protein-coding and non-coding sequences of mRNA. In the cytoplasm, this includes regulatory events that dictate which mRNAs are preferentially degraded or translated to make proteins. Together, these regulatory processes specify the identity and abundance of proteins present in each cell, and consequently, cell properties. In order for cells to progress through different stages of development, its mRNAs undergo extensive stage-specific regulation. Defects in mRNA regulation are the direct cause of many human diseases, thus an understanding of how mRNAs are regulated is essential. In the proposed study, we will comprehensively characterize mRNA regulatory events that drive cells through different stages of male germ cell development. This cellular program depends on undefined mRNA regulatory programs. Our plan is to isolate postnatal mouse germ cells at different steps in this pathway and use transcriptome-wide tools to 1) identify changes in mRNA expression and translation, 2) understand the functional significance of these changes, and 3) determine how these changes are controlled at the molecular level. We will use an approach established in our laboratory that combines dual fluorescence cell labeling in transgenic mice and fluorescence activated cell sorting to isolate cells at different stages of development. Combining this approach with deep sequencing, biochemical, and bioinformatic tools, we will reveal multiple layers of mRNA-based gene control during germ cell development. Altogether, we will gain important insights into the functions and mechanisms of mRNA regulation in mammalian cell development in unprecedented molecular and cellular detail. As a result, this study will lead to an improved understanding of how gene expression is controlled through mRNA regulation during germ cell development. The data has the potential to provide important new insights into human reproductive biology and fertility. This study will also reveal molecular mechanisms controlling mammalian cell differentiation and proliferation, and therefore will be of direct relevance to human development and disease.

项目摘要 拟议工作的长期目标是更好地了解基因表达的调节 在哺乳动物细胞发育过程中。该提案重点是研究基因的转录后控制 表达，从单个基因制成的mRNA可以以不同的方式改变基因 输出。在细胞核中，这包括改变mRNA的蛋白质编码和非编码序列。在 细胞质，这包括调节事件，这些事件决定了哪些mRNA优先降解或翻译 制作蛋白质。这些调节过程共同指定了存在的蛋白质的身份和丰度 在每个细胞中，因此是单元格性质。为了使细胞通过不同的阶段进行 开发，其mRNA受到广泛的特定阶段调节。 mRNA调节中的缺陷是 许多人类疾病的直接原因，因此对MRNA如何调节的理解至关重要。 在拟议的研究中，我们将全面地表征mRNA调节事件，这些事件驱动细胞通过 男性生殖细胞发育的不同阶段。该细胞程序取决于未定义的mRNA调节 程序。我们的计划是在此途径的不同步骤中分离出产后小鼠生殖细胞并使用 整个转录组的工具1）确定mRNA表达和翻译的变化，2）了解 这些变化的功能意义，以及3）确定如何在分子上控制这些变化 等级。我们将使用实验室中建立的一种方法，该方法结合了双荧光细胞标记 转基因小鼠和荧光激活的细胞分类以在不同发育阶段分离细胞。 将这种方法与深层测序，生化和生物信息学工具相结合，我们将揭示多个 生殖细胞发育过程中基于mRNA的基因对照层。 总之，我们将获得对mRNA调节功能和机制的重要见解 哺乳动物细胞发育以前所未有的分子和细胞细节。结果，这项研究将导致 对在生殖细胞期间通过mRNA调节控制基因表达的理解有了改进的了解 发展。数据有可能为人类生殖生物学提供重要的新见解，并 生育能力。这项研究还将揭示控制哺乳动物细胞分化和的分子机制 扩散，因此将与人类发展和疾病直接相关。