Novel optogenetic method for control of protein synthesis

控制蛋白质合成的新型光遗传学方法

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要局部蛋白质合成允许蛋白质在现场和点播的所有快速表达从神经元中运输前脱蛋白。将基因表达限制为可以来自细胞体的个体结构的手段，例如突触和生长锥。但是，尽管局部翻译对正常功能的重要性，但最广泛使用的方法是通过低空间或时间分辨率来操纵蛋白质的合成该建议我们将生成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不同的mRNA 和细胞类型。第二目的产生了一种编译功能的方法来急性刺激局部蛋白质合成。 ASA原理证明，我们将使用剧院方法来确定局部β-的需求和足够性肌动蛋白翻译用于响应神经元活性的原发性海马神经元的突触可塑性。肌动蛋白mRNA迅速在树突中迅速转运到其被翻译的活性突触中，纳斯特β- 肌动蛋白蛋白专门用活性树突脊柱保留。这将为研究人员在特定的转录后基因调节中获得精致的控制亚细胞室。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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.