中文摘要
本项目致力于建立非编码RNA检测与标记的新技术和新方法。为了研究新非编码RNA的作用机理,我们将发展RNA交联技术研究RNA/蛋白质双色作用网络,并发展新的RNA活体成像的方法研究非编码RNA的转运和定位机制。我们发展了适用于捕获蛋白质/RNA及RNA/RNA瞬态相互作用的,可切断的交联探针。我们发展了活细胞内特定RNA特定位点的共价标记新方法,这将能够使得我们在引入最小干扰的情况下,对目标RNA进行高精度的荧光、核磁、顺磁、红外标记。以上方法在国际上未见报导,均是我们首创。在此项目的支持下,我们将为本重大研究计划所有课题组提供必不可少的通用工具箱,并在RNA跨核膜转运机理研究方面获得突破。
英文摘要
In this project, we will focus to develop new technologies and new methods for non-coding RNA (ncRNA) detection and labelling. In order to investigate the functions of these new ncRNA, we will develop novel RNA crosslinking methods to discover dynamic functional RNA/protein interactions and RNA/RNA interactions. We will develop novel sequence and site specific labeling methods to incorporate azide and alkyne probes into RNA in living cells. In conjunction with novel bioorthogonal reactions, we will be able to image the localization and transportation of the low abundance ncRNA in mammalian cells. Through this method, we will also be able to introduce NMR, IR and EPR probes site-specifically into RNA. The crosslinking and imaging methods we will develop form an essential and integral ncRNA research toolbox, which will help all of the nRNA research community. These methods will significantly advance our ability to investigate RNA structure and function, both in vivo and in vitro.
结题摘要
本项目致力于建立非编码RNA检测与标记的新技术和新方法。为了研究新非编码RNA的作用机理,我们发展了RNA交联技术研究RNA/蛋白质双色作用网络,并发展新的RNA活体成像的方法研究非编码RNA的定位机制。我们发展了适用于捕获蛋白质/RNA及RNA/RNA瞬态相互作用的交联探针。我们发展了活细胞内特定RNA特定位点的共价标记新方法,这将能够使得我们在引入最小干扰的情况下,对目标RNA进行高精度的荧光、核磁、顺磁、红外标记。以上方法在国际上未见报导,均是我们首创。在此项目的支持下,我们为本重大研究计划所有课题组提供必不可少的通用工具箱。同时本项目阶段性成果在Angew. Chem. Intl. Ed,PNAS,Discovery Medicine和Chembiochem等杂志上发表文章4篇,并申请中国专利1项。
