Quantitative mapping of synaptic proteins and mRNA in neurons & a nano-analytical platform to probe binding mechanisms of RNA-binding proteins (A08)

神经元突触蛋白和 mRNA 的定量图谱

• 批准号: 249370415
• 金额: --
• 依托单位: Max-Planck-Institut für Hirnforschung
• 依托单位国家: 德国
• 项目类别: Collaborative Research Centres
• 财政年份: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/249370415?language=en
• 关键词: Quantitative; mapping; synaptic; proteins; mRNA

Synaptic activity is coupled to protein copy numbers, which vary and are regulated by the presence of mRNA and miRNA. We will develop a nano-scale imaging platform that will allow a comprehensive analysis of protein, mRNA and microRNA “localizome”, by visualizing multiple proteins and mRNA species in the same cell using single-molecule super-resolution imaging with large multiplexing capabilities. Next to localizing and quantifying protein and mRNA, we aim to develop tools that allow super resolution of molecules that regulate miRNA/mRNA, and newly transcribed proteins. In addition, we will develop a nano-analytical platform to study the mechanism of RNA-binding proteins binding to their target RNA sequence. This novel platform will allow the study of unmodified proteins, and provide information on binding equilibria and kinetics, sequence specificity, interaction with co-factors and the influence of mutations

突触活动与蛋白质拷贝数相关，蛋白质拷贝数因mRNA和miRNA的存在而异并受其调节。我们将开发一个纳米级的成像平台，通过使用具有大复用能力的单分子超分辨率成像来可视化同一细胞中的多个蛋白质和mRNA种类，从而对蛋白质、mRNA和microRNA“localizome”进行全面分析。接下来定位和定量蛋白质和mRNA，我们的目标是开发工具，允许超分辨率的分子调节miRNA/mRNA，和新转录的蛋白质。此外，我们将开发一个纳米分析平台，以研究RNA结合蛋白与其靶RNA序列结合的机制。这个新的平台将允许研究未修饰的蛋白质，并提供关于结合平衡和动力学、序列特异性、与辅因子的相互作用以及突变影响的信息