Ptbp2 function and global mRNA regulation during mammalian cell development

哺乳动物细胞发育过程中 Ptbp2 功能和全局 mRNA 调控

• 批准号: 8728294
• 负责人: Donny D Licatalosi
• 金额: $ 30.12万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8728294
• 关键词: Alleles; Animals; Biochemical; Bioinformatics; Cell Separation; Cells; Contraceptive methods; Data; Data Set; Defect; Development; Event; Extensive Stage; Flow Cytometry; Functional RNA; Gametogenesis; Gene Expression; Gene Expression Profile; Genes; Genetic; Genetic Models; Germ Cells; Goals; Human; Lead; Life; Male Contraceptions; Mammalian Cell; Mammals; Messenger RNA; Methods; Molecular; Mus; Null Lymphocytes; Phase; Polyadenylation; Population; Post-Transcriptional Regulation; Process; Proteins; RNA; RNA Splicing; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Reproductive Biology; Research; Resolution; Ribosomal Proteins; Ribosomes; Role; Spermatogenesis; Staging; System; Technology; Therapeutic; Tissues; Transgenic Mice; Wild Type Mouse; Work; abstracting; assisted reproduction; base; cell type; deep sequencing; human disease; improved; in vivo; in vivo Model; insight; male; mouse development; novel strategies; public health relevance; research study; sperm cell; tool; transcriptome sequencing

DESCRIPTION (provided by applicant): PROJECT SUMMARY/ ABSTRACT 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 mRNA, which can be regulated in many ways to provide multiple layers of gene expression control. In order for cells to progress through different stages of development, its mRNAs must undergo extensive stage-specific changes. Defects in mRNA regulation are the direct cause of many human diseases, thus a better understanding of how mRNAs are regulated is essential. Our plan is to isolate mouse cells that are in specific stages of development and perform comprehensive analyses to identify changes in mRNA regulation, understand the functional significance of these changes, and determine how these changes are controlled at the molecular level. To do so, we have developed a new approach to purify male mouse germ cells in each of the major stages of gametogenesis (the process by which sperm are generated). Combining this new cell isolation strategy with powerful genetic, biochemical, deep sequencing, and bioinformatic tools, we will be able to perform comprehensive, global analyses of multiple layers of mRNA-based gene control in wild type mice as well as mice lacking an essential protein necessary for proper RNA regulation. In doing so, we will gain important insights into mechanisms and functions of mRNA regulation in different stages of 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 cell development, and has the potential to provide important new insights into human reproductive biology and the molecular mechanisms controlling fertility.

描述（由申请人提供）： 项目摘要/摘要拟议的工作的长期目标是更好地了解在哺乳动物细胞发育过程中如何调节基因表达。该建议的重点是对mRNA的研究，mRNA可以通过多种方式调节，以提供多个基因表达控制层。为了使细胞通过不同的阶段发展 在发展中，其mRNA必须经历广泛的特定阶段特定变化。 mRNA调控缺陷是许多人类疾病的直接原因，因此对mRNA的调节方式有了更好的了解。我们的计划是隔离处于发育特定阶段的小鼠细胞并进行全面的分析以识别mRNA调节的变化，了解这些变化的功能意义，并确定如何在分子水平控制这些变化。为此，我们开发了一种新的方法来净化男配子发生的每个主要阶段（生成精子的过程）。将这种新的细胞隔离策略与强大的遗传，生化，深层测序和生物信息学工具相结合，我们将能够对野生型小鼠中多层基于mRNA的基因控制的多层分析以及缺乏适当RNA调节所必需的必要蛋白质的小鼠进行全面的全球分析。在此过程中，我们将以前所未有的分子和细胞细节的不同阶段的不同阶段中对mRNA调节的机制和功能的重要见解。结果，这项研究将提高人们对细胞发育过程中如何通过mRNA调节来控制基因表达的理解，并有潜力为人类生殖生物学和控制生育能力的分子机制提供重要的新见解。