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：
8642412
负责人：
Gilad Barnea
金额：
$ 24.05万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
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/8642412
关键词：
Affect Afferent Neurons Alleles Animal Model Architecture Binding Biochemical Biological Assay Brain Cataloging 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 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 Optics Pattern Population Positioning Attribute Post-Translational Protein Processing Property Protein Binding Proteins Receptor Gene Regulation Resolution Rest Seminal Specificity System Technology Tertiary Protein Structure Therapeutic Intervention Time Tissues Transcription Coactivator Variant Viral base cell type combinatorial design emerin histone modification human disease in vivo innovation lamin B receptor millisecond novel olfactory receptor programs promoter public health relevance research study spatiotemporal synthetic construct tool 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凌驾于DNA编码的调控信息之上，这为使用表观遗传修饰剂稳定地改变体内基因表达程序提供了令人兴奋的机会。然而，这种愿望受到了我们在时间调控时尚中改变受限细胞类型中特定靶基因表观遗传状态的能力的限制的挑战。因此，我们建议将能够提供严格的体内时空控制的新颖的遗传方法与能够以序列特异性方式靶向特定基因组位点的创新生化工具相结合。我们将修改我们之前为特定神经元群的诱导标记而设计的检测方法，称为 Tango，以控制合成 TALE（转录激活因子样效应器）融合蛋白的表达，该融合蛋白将与目标基因组位点结合并改变其表观遗传特性。作为这些原理验证实验的模型，我们将使用遗传、表观遗传和生物化学上易于处理的小鼠嗅觉系统。正如我们之前所表明的，嗅觉感觉神经元（OSN）中嗅觉受体（OR）基因的单基因和单等位基因表达受到表观遗传调控，无论是在翻译后组蛋白修饰水平上，还是在核组织和活性和分布的水平上。沉默或等位基因。因此，我们建议使用 TANGO 系统的变体在特定 OSN 亚群中以诱导方式表达对 OR 基因及其调节增强子具有特异性的 TALE 融合蛋白。通过这种方式，我们将改变活性或沉默 OR 的表观遗传状态，并诱导它们重新定位到不同的核区域，以稳定改变它们的表达模式。这种化学或光学控制的表观遗传操作策略将直接适用于小鼠中的任何其他细胞类型，并且与病毒递送方法兼容，这将使我们的方法适用于未来人类疾病的治疗干预。