Development of aptamere based RNA labeling techniques for analysis of RNA dynamics inside living cells via FRET

许多细胞过程受到不同种类RNA的调控和影响，如非编码RNA、mRNA和毒性RNA。为了详细了解这些过程，为了研究RNA和RNA与其他生物大分子的相互作用，一种能够在细胞内进行实时分析的成像方法是必不可少的。Jaffrey实验室最近开发了菠菜适体，可以标记和研究活细胞内的RNA。菠菜适体代表了开发绿色荧光RNA适体标签的第一种方法。进一步的研究和开发是必要的，以创造一系列技术，类似于荧光蛋白，允许研究RNA-RNA或rna -蛋白质相互作用。因此，最重要的是对活细胞内RNA及其相互作用伙伴进行直接观察的可能性。在Jaffrey教授小组的研究项目中，我将开发(1)菠菜适体的新的红色荧光变体，它模仿红色荧光蛋白，并将可用的适体标签扩展到红光光谱。基于这个新的红移适体，我将开始建立(2)荧光蛋白标签和RNA适体标签之间的FRET-System。该系统将用于通过荧光显微镜研究活细胞内蛋白质- rna相互作用。

Many cellular processes are regulated and influenced by different RNA species, like non-coding RNA, mRNA and toxic RNA. For a detailed understanding of these processes and for investigation of RNA and RNA interaction with other biomacromolecules, an imaging approach that allows real-time analysis inside cells is essential. The Jaffrey lab recently developed the spinach aptamer which enables to label and study RNA inside living cells. The spinach aptamere represents the first approach to develop a green fluorescent RNA aptamer tag. Further research and development is necessary to create a repertoire of techniques, similar to fluorescent proteins, that allow the study of RNA-RNA or RNA-protein interactions. Most important is hereby the possibility of performing direct observation of RNA and its interaction partners inside living cells.During my research project in the group of Prof. Jaffrey I will develop (1) new red fluorescent variant of the spinach aptamer that mimics red fluorescent proteins and expands the repertoire of available aptamer tags towards the red light spectrum. Based on this new red shifted aptamer I will start to establish (2) a FRET-System between fluorescent protein tags and RNA aptamer tags. This system will be used to study protein-RNA interaction inside living cells by fluorescence microscopy.