RNA研究是当前生命科学研究的热点。随着高通量测序技术的发展，RNA的研究进入了转录组学时代。同样RNA研究中的难点mRNA二级结构同样也需要从转录组进行整体分析。本项目拟基于识别RNA高级结构的分子探针建立活体mRNA二级结构的高通量测序技术。主要研究内容为 (1) 设计合成识别RNA二级结构的分子探针。对探针进行衍生功能化，进行叠氮修饰和光诱导基团保护来降低测序背景噪音和扩大技术应用范围；(2) 以已知结构的5S rRNA为模型，通过逆转录终止实验来验证分子探针在体外和体内识别RNA二级结构的准确度。经验证后再将分子探针用于mRNA的二级结构的测定；(3) mRNA经分子探针修饰后进行建库、测序分析，并将技术应用于一些生物医学问题研究。目前mRNA的高通量测序在国际上正处于起步阶段，该项目的实施将有助于理解mRNA结构在调控基因表达和基因组稳定性中发挥的多重功能。

The research of RNA is the hot spot in biological field. With the development of Next generation high-throughput sequencing technology, people can study RNA in transcriptome. The secondary structure of mRNA, as the biggest challenge of RNA research, also need be studied in transcriptome. So our project will develop a high-throughput sequencing technology to mRNA structure. We will: (1) design and synthesis the molecular probes for RNA secondary structure. And we also modify the probes with azide and photo-caged groups, which can reduce the background noise and increase the application of this technology; (2) Based on the structure data of 5S rRNA, we will firstly study the structure recognition of probes to 5S rRNA by primer extension stop experiments in vitro and in vivo, which can verify the accuracy of the probes. Then the probes will apply with the mRNA secondary structure research. (3) The library construction and sequencing of mRNA after the molecular probes treatment. We will also use this technology to study some important biological and medical issues. Now the high-throughput of mRNA structure is only starting the stage. The accomplishment of this project has great significance to understand the function of mRNA secondary structure in gene regulation and gene stability.