Novel optogenetic method for control of protein synthesis

控制蛋白质合成的新型光遗传学方法

• 批准号: 10369685
• 负责人: Kenneth Myers
• 金额: $ 19.56万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-15 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369685
• 关键词: Acute; Address; Affect; Amyotrophic Lateral Sclerosis; Binding; Binding Proteins; Cells; Code; Cognition Disorders; Complex; Data; Defect; Dendrites; Dendritic Spines; Development; Disease; Etiology; Excision; Fluorescent in Situ Hybridization; Fragile X Syndrome; Gene Expression; Generations; Genetic; Genetic Translation; Goals; Growth Cones; Hippocampus (Brain); Human; Individual; Label; Laboratories; Learning; Light; Link; Memory; Messenger RNA; Methodology; Methods; Morphology; Neurodevelopmental Disorder; Neurons; Organism; Pharmacology; Photosensitizing Agents; Play; Post-Transcriptional Regulation; Protein Biosynthesis; Proteins; RNA-Binding Proteins; Reactive Oxygen Species; Reporter; Research Personnel; Resolution; Role; Site; Small RNA; Specificity; Spinal Muscular Atrophy; Structure; Synapses; Synaptic plasticity; Techniques; Testing; Transcript; Translating; Translations; Vertebral column; Visualization; autism spectrum disorder; axon growth; axon guidance; beta Actin; cell type; chromophore; design; efficacy evaluation; experimental study; gain of function; high resolution imaging; imaging modality; improved; inhibitor; loss of function; migration; mutant; nanoscale; nervous system disorder; neuron development; novel; optogenetics; postsynaptic; prevent; protein expression; response; spatiotemporal; submicron; supernova; technique development; temporal measurement; tool

Project Summary Local protein synthesis allows for the rapid expression of proteins on-site and on-demand, as opposed to transporting pre-existing proteins from elsewhere. In neurons, the site-specific translation of mRNAs provides a means for restricting gene expression to individual structures that can be far from the cell body, such as synapses and growth cones. Dysregulated protein synthesis is implicated in a variety of neurodevelopmental and cognitive disorders. However, despite the importance of local translation for normal neuronal function, the most widely used methods for manipulating protein synthesis suffer from low spatial or temporal resolution. In this proposal we will generate new optogenetic methods for regulating the local translation of individual mRNA molecules. These methods will be broadly applicable to different mRNAs for use in a wide array of organisms and cell types. The first aim develops a new loss-of-function approach to inhibit local translation, while the second aim generates a complementary gain-of-function method to acutely stimulate local protein synthesis. As a proof of principle, we will use these methods to determine both the requirement and sufficiency of local β- actin translation for synaptic plasticity using primary hippocampal neurons. In response to neuronal activity, β- actin mRNA is rapidly transported within dendrites to active synapses where it is translated, and the nascent β- actin protein is specifically retained within the active dendritic spine. In this proposal, we will develop new tools that will provide researchers with exquisite control over post-transcriptional gene regulation in specific subcellular compartments.

项目概要 局部蛋白质合成允许蛋白质在现场和按需快速表达，而不是 从其他地方运输预先存在的蛋白质。在神经元中，mRNA 的位点特异性翻译提供了 将基因表达限制在远离细胞体的个体结构的方法，例如 突触和生长锥。蛋白质合成失调与多种神经发育有关 和认知障碍。然而，尽管局部翻译对于正常神经元功能很重要， 最广泛使用的操纵蛋白质合成的方法都存在空间或时间分辨率低的问题。在 根据该提案，我们将产生新的光遗传学方法来调节单个 mRNA 的局部翻译 分子。这些方法将广泛适用于多种生物体中的不同 mRNA 和细胞类型。第一个目标是开发一种新的功能丧失方法来抑制本地翻译，而 第二个目标是产生一种互补的功能获得方法来急剧刺激局部蛋白质合成。 作为原理证明，我们将使用这些方法来确定局部 β- 的要求和充分性 使用初级海马神经元进行肌动蛋白翻译以实现突触可塑性。响应神经元活动，β- 肌动蛋白 mRNA 在树突内快速转运到活跃的突触，并在那里进行翻译，新生的 β- 肌动蛋白特异地保留在活跃的树突棘内。在这个提案中，我们将开发新的工具 这将为研究人员提供对特定转录后基因调控的精确控制 亚细胞区室。

项目成果

Actin capping protein regulates postsynaptic spine development through CPI-motif interactions.

• DOI: 10.3389/fnmol.2022.1020949
• 发表时间: 2022
• 期刊: FRONTIERS IN MOLECULAR NEUROSCIENCE
• 影响因子: 4.8
• 作者: Myers, Kenneth R.;Fan, Yanjie;McConnell, Patrick;Cooper, John A.;Zheng, James Q.
• 通讯作者: Zheng, James Q.