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

突触活性与蛋白质拷贝数有关，蛋白质拷贝数随蛋白质拷贝数的变化而变化，并受mRNA和miRNA的存在调节。我们将开发一个纳米级成像平台，通过使用具有强大多路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信使核糖核酸物种，从而实现对蛋白质、信使核糖核酸和微核糖核糖核酸体的全面分析。除了定位和量化蛋白质和信使核糖核酸，我们的目标是开发能够超分辨调节miRNA/信使核糖核酸的分子和新转录的蛋白质的工具。此外，我们将开发一个纳米分析平台来研究RNA结合蛋白与其目标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、与辅助因子的相互作用以及突变的影响的信息

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