Temporal, cell type- and locus-specific epigenetic control in transgenic mice

转基因小鼠的时间、细胞类型和位点特异性表观遗传控制

• 批准号: 8735923
• 负责人: Linda Madisen
• 金额: $ 46.36万
• 依托单位: ALLEN INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8735923
• 关键词: Adult; Animals; Binding; Biological Assay; Biological Models; Biology; Brain; Brain Diseases; Cell Maintenance; Cell physiology; Cells; Cellular Morphology; Development; Disease; Doxycycline; EP300 gene; ES Cell Line; Ensure; Epigenetic Process; Future; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genome; HDAC1 gene; Histocompatibility Testing; In Vitro; Knock-in Mouse; Label; Life; LoxP-flanked allele; Malignant Neoplasms; Mediating; Mental disorders; Methodology; Modeling; Modification; Molecular; Mus; Neocortex; Neurons; Organ; Organism; Phenotype; Proteins; Reporter; Response Elements; Specificity; Technology; Testing; Tetracyclines; Time; Tissues; Trans-Activators; Transcription Coactivator; Transgenes; Transgenic Mice; Transgenic Organisms; Validation; Withdrawal; Work; base; cell type; design; design and construction; embryo cell; embryonic stem cell; human ZNF45 protein; in vivo; insight; interest; knock-down; mouse genome; nerve stem cell; overexpression; prevent; progenitor; public health relevance; recombinase; small hairpin RNA; tissue culture; tool; transgene expression

DESCRIPTION (provided by applicant): Mammalian organs are built of a variety of intermingled cell types, the full extent and diversity of which are unknown. Specification and maintenance of cell identity, from a single cell embryo to any terminally differentiated cell such as an adult neuron, rely heavily on epigenetic mechanisms that act upon the genome to enable or prevent expression of specific sets of genes. In doing so, epigenetic regulation of gene expression enables a single genome to encode a diverse array of cell types in a multicellular organism. Epigenetic modifications ensure that a cell proceeds through a limited set of gene expression possibilities, and once differentiated, that a cell maintains its phenotype throughout its lifetime. As a consequence, disruption of epigenetic modification can result in a variety of diseases, including neurodevelopmental and psychiatric diseases, and cancer. We propose to develop a set of mouse transgenic tools that have the potential to transform the way epigenetic modifications can be studied and understood in this mammalian model system. The new tools will provide regulated expression of epigenetic modifiers in a variety of cell types and their progenitors, while permanently marking these cells or their progeny for examination at any later time point. Moreover, we will develop technology, in which a single locus in the mouse genome can be subjected to epigenetic perturbations in specific cells and at specific times in the animal's life. We will employ these tools to gain understanding of cell-type identity in the cortex as well as the mechanisms responsible for generating and maintaining this identity. Epigenetic regulators have been implicated in a number of brain diseases, and the tools we are developing will facilitate modeling or testing specific hypotheses that relate to perturbations of epigenetic phenomena. The insights gained may also provide guidance for the generation of specific cell types in vitro, and directions for 'repurposing' certain cell types in vivo. Although the focus of ur studies will be on the mouse brain, the versatile and modular design of our tools will enable their use in studying development, function or disease in any other tissue or cell type. We will ensure easy access to all tools we generate to maximize their impact on understanding various aspects of mammalian biology.

描述(申请人提供)：哺乳动物的器官由各种混合的细胞类型组成，其完整的范围和多样性尚不清楚。从单细胞胚胎到任何终末分化的细胞，如成年神经元，细胞特性的确定和维持在很大程度上依赖于作用于基因组的表观遗传机制，以启用或阻止特定组基因的表达。在这样做的过程中，基因表达的表观遗传调节使单个基因组能够编码多细胞生物体中不同类型的细胞。表观遗传修饰确保细胞通过一组有限的基因表达可能性，一旦分化，细胞在其一生中保持其表型。因此，表观遗传修饰的中断可能导致各种疾病，包括神经发育和精神疾病，以及癌症。我们建议开发一套小鼠转基因工具，这些工具有可能改变在这个哺乳动物模型系统中研究和理解表观遗传修饰的方式。新工具将在各种类型的细胞及其祖细胞中提供表观遗传修饰物的调控表达，同时永久标记这些细胞或其后代，以便在以后的任何时间点进行检查。此外，我们将开发一种技术，在这种技术中，小鼠基因组中的单个基因座可以在动物生命中的特定细胞和特定时间受到表观遗传扰动。我们将使用这些工具来了解大脑皮层的细胞类型身份，以及负责产生和维持这种身份的机制。表观遗传调节器已经与许多大脑疾病有关，我们正在开发的工具将有助于建模或测试与表观遗传现象的扰动有关的特定假说。所获得的洞察力也可能为体外产生特定类型的细胞提供指导，并为在体内重新定位某些类型的细胞提供方向。尽管我们的研究重点将放在小鼠的大脑上，但我们工具的多功能性和模块化设计将使他们的 用于研究任何其他组织或细胞类型的发育、功能或疾病。我们将确保方便地使用我们生成的所有工具，以最大限度地发挥它们对理解哺乳动物生物学各个方面的影响。