权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

AN INDUCIBLE MOLECULAR MEMORY SYSTEM TO RECORD TRANSIENT STATES OF CNS CELLS

记录中枢神经系统细胞瞬态的可诱导分子记忆系统

基本信息

批准号：
9301354
负责人：
Robi D Mitra
金额：
$ 121.36万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-18 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9301354
关键词：
Alpha Cell Alzheimer&apos s Disease Amygdaloid structure Anatomy Animal Behavior Behavior Behavioral Binding Brain Cells Central Nervous System Diseases ChIP-seq Chemicals Communities Complex DNA-Binding Proteins DNA-Protein Interaction Data Development Disease Embryo Environment Epigenetic Process Event FOS gene Genes Genetic Genetic Transcription Genome Genome Mappings Genomics Huntington Disease Immediate-Early Genes In Vitro Informatics Investigation Knowledge Lateral Learning Maps Mediator of activation protein Memory Methods Molecular Molecular Analysis Mus Nerve Degeneration Neurobiology Neurodegenerative Disorders Neurons Organ Organism Outcome Parkinson Disease Phenotype Physiological Population Reagent Reporter Reproducibility Resistance Role Stem cells Stimulus Structure System Techniques Technology Testing Time Transposase Ursidae Family Vertebrates Viral brain cell candidate identification cell type cognitive process enhancer binding protein experimental study fear memory genome-wide in vivo memory consolidation molecular phenotype nerve stem cell neural circuit neuronal circuitry new technology novel progenitor protein E public health relevance relating to nervous system response tool transcription factor transcriptome sequencing vocalization

项目摘要

DESCRIPTION (provided by applicant): The brain is a remarkably complex organ comprised of hundreds of unique cell types that are organized to form sophisticated neural circuits. Although we have made progress toward understanding brain function and development, it is clear there is still much to be learned. Currently, all genome-wide methods that could be brought to bear on functional studies of the brain are destructive, meaning that as a genomic analysis is performed on a population of cells, the cells are destroyed. This fact limits our ability to connec early molecular events in the cells of the brain with later behavioral or cellular changes. For example, it is currently impossible to connect transcriptional changes in a neuron with knowledge of whether or not the cell was successfully incorporated into a memory trace. Similarly, it is not feasible to connect the early molecular events that occur in a neuronal progenitor cell with the final cell fate decision made by the cell. We have set out to develop a transformative technology that can record molecular events at the time that they occur and can then be read out later after any defined period of time. We have a novel technology called transposon `Calling Cards' that, in culture, provides cells with a molecular memory of protein-DNA interactions that occur at a particular moment in time. Here, we propose to adapt this technology for use in vivo enabling a retrospective genomic analysis of molecular events. We will demonstrate the utility of this technology by completing four test-case experiments that cannot be done with existing methods. Specifically, we will test the method by: 1) retrospectively identifying candidate transcription factors that control the specification of cell types in the CNS 2) identifying features that distinguish neurons resistant to neurodegeneration in vivo, 3) identifying the neurons that become active during mouse vocalization behavior while simultaneously mapping the genome-wide binding of activity-dependent transcription factors in these neurons, and 4) identifying the molecular features that distinguish neurons that were incorporated into a fear memory trace from those that were not incorporated.

描述(申请人提供)：大脑是一个非常复杂的器官，由数百种独特的细胞类型组成，这些细胞被组织起来形成复杂的神经电路。尽管我们在了解大脑功能和发育方面取得了进展，但很明显，仍有很多东西需要学习。目前，所有可以用于大脑功能研究的全基因组方法都是破坏性的，这意味着当对一群细胞进行基因组分析时，这些细胞就会被摧毁。这一事实限制了我们将大脑细胞中的早期分子事件与后来的行为或细胞变化联系起来的能力。例如，目前还不可能将神经元的转录变化与细胞是否成功整合到记忆痕迹中联系起来。同样，将神经元前体细胞中发生的早期分子事件与细胞做出的最终细胞命运决定联系起来也是不可行的。我们已经着手开发一种变革性的技术，它可以在分子事件发生时记录它们，然后在任何确定的时间段后被读出。我们有一种名为转座子‘通话卡’的新技术，在培养中，它为细胞提供了在特定时刻发生的蛋白质-DNA相互作用的分子记忆。在这里，我们建议将这项技术应用于体内，以实现对分子事件的追溯基因组分析。我们将通过完成四个用现有方法无法完成的测试用例实验来演示这项技术的实用性。具体地说，我们将通过以下方式对该方法进行测试：1)追溯鉴定控制CNS中细胞类型规范的候选转录因子；2)鉴定区分体内抗神经变性神经元的特征；3)鉴定在小鼠发声行为中变得活跃的神经元，同时映射这些神经元中活性依赖的转录因子的全基因组结合；以及4)鉴定区分纳入恐惧记忆痕迹的神经元与未纳入恐惧记忆痕迹的神经元的分子特征。