Controlling epigenetic states and nuclear architecture in the brain

控制大脑中的表观遗传状态和核结构

基本信息

批准号：
9275951
负责人：
Gilad Barnea
金额：
$ 22.73万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-15 至 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9275951
关键词：
ARNT gene Affect Alleles Animal Model Architecture Binding Binding Proteins Biochemical Biological Assay Brain Catalogs Chemicals Chimeric Proteins Chromatin DNA DNA Binding Domain DNA Sequence Development Dimerization Distant Elements Enhancers Epigenetic Process Future Gene Expression Gene Expression Regulation Gene Order Gene Targeting Genes Genetic Transcription Genomics Goals Histones Label Light Location Mediating Methods Modeling Modification Mus Names Nervous System Physiology Neuraxis Neurodevelopmental Disorder Neurons Nuclear Nuclear Envelope Nuclear Lamina Olfactory Pathways Optics Pattern Population Positioning Attribute Post-Translational Protein Processing Property Proteins Receptor Gene Resolution Rest Seminal Specificity System Technology Tertiary Protein Structure Therapeutic Intervention Time Tissues Transcription Coactivator Variant Viral base biochemical tools cell type combinatorial design dimer emerin epigenetic regulation experimental study genetic approach genetic information histone modification human disease in vivo innovation lamin B receptor millisecond novel olfactory receptor olfactory sensory neurons programs promoter public health relevance spatiotemporal synthetic construct temporal measurement transcription factor

项目摘要

DESCRIPTION (provided by applicant): The realization that epigenetic control of gene expression can override regulatory information encoded in DNA provides the exciting opportunity to stably alter gene expression programs in vivo with the use of epigenetic modifiers. However, this aspiration is challenged by limitations in our ability to alter the epigenetic state of specific target genes in restricted cell types in a temporally regulated fashio. For this reason, we propose to combine novel genetic approaches that afford tight spatiotemporal control in vivo with innovative biochemical tools that allow the targeting of specific genomic loci in a sequence-specific manner. We will modify an assay that we previously designed for the inducible labeling of specific neuronal populations, named Tango, towards the controlled expression of synthetic TALE (Transcription Activator Like Effectors)-fusion proteins that will bind to target genomic loci and alter their epigenetic properties. As a model for these proof-of-principle experiments we will use the genetically, epigenetically and biochemically tractable mouse olfactory system. As we previously showed, the monogenic and monoallelic expression of olfactory receptor (OR) genes in olfactory sensory neurons (OSNs) is epigenetically regulated, both at the level of post-translational histone modifications and at the level of nuclear organization and distribution of active and silent OR alleles. Therefore, we propose to express TALE-fusion proteins with specificity for OR genes and their regulating enhancers in an inducible fashion in specific OSN subpopulations using variations of the TANGO system. This way we will alter the epigenetic state of active or silent ORs, and induce their re-positioning to distinct nuclear territories with the goal of stably altering their expresson pattern. This strategy of chemically or optically controlled epigenetic manipulations will be directly applicable to any other cell type in the mouse, and compatible with viral delivery methods that will make our approach applicable to future therapeutic interventions for human disease.

描述（由申请人提供）：认识到，基因表达的表观遗传控制可以覆盖DNA中编码的调节信息，这为使用表观遗传修饰符在体内稳定改变基因表达程序提供了令人兴奋的机会。但是，这种攻击受到我们改变时间调节的fashio细胞类型中特定靶基因表观遗传状态的能力的局限性的挑战。因此，我们建议将新型的遗传方法与创新的生化工具相结合，这些方法可以在体内进行严格的时空控制，允许以序列特异性方式靶向特定的基因组基因局。我们将修改我们先前设计的用于针对特定神经元种群的可诱导标记，称为Tango，朝着合成故事的受控表达（转录激活剂（例如效应子）） - 融合蛋白，该蛋白将与靶基因组基因群结合并改变其表观遗传特性。作为这些原则实验的模型，我们将使用遗传，表观遗传和生化的小鼠嗅觉系统使用该模型。如前所述，嗅觉受体基因（OR）基因（OSN）的单相表达（OSN）在遗传学后均受到表观调节，无论是在翻译后组蛋白修饰的水平上，以及在活动的核组织和分布和活性和静音或静音或静音或静音或静音的水平下。因此，我们建议使用探戈系统的变化在特定的OSN亚群中以特异性或基因的特异性表达具有特异性或基因及其调节增强子的故事融合蛋白。这样，我们将改变活跃或沉默的OR的表观遗传状态，并诱导其重新定位到不同的核领土，以稳定地改变其表达模式。这种化学或光学控制的表观遗传操作的策略将直接适用于小鼠中的任何其他细胞类型，并与病毒递送方法兼容，这将使我们的方法适用于未来对人类疾病的治疗干预措施。