基于核酸纳米结构信号放大元件的RNA可变剪接测定及原位荧光成像研究

RNA alternative splicing is the important mechanism for the regulation of gene expression and protein diversity. It is also closely related to the occurrence and development of cancer and other disease. The mis-splicing induced variants are the potential biomarkers for cancer diagnosis and prognosis. This project focuses on the construction of methods for the highly sensitive and specific detection and in-situ fluorescence imaging of RNA alternative splicing. Then we will further explore the correlations between cancer and expression difference of variants. In this project, we plan to study the construction of nucleic acid nanostructures which can be used as the signal amplification elements. Combining with the structure change of variant molecules induced by RNA alternative splicing, we attempt to design the highly specific signal transition and amplification strategies for monitoring RNA alternative splicing variants. After that, we will develop some novel methods for sensitive and selective detection of RNA alternative splicing events and in-situ fluorescence imaging of single variants molecule in cells. Finally, we will investigate the expression and distribution differences of splicing variants in lung cancer cells, and explore the relationship between these differences and cancers. These studies will provide valuable theoretical references and technical supports for cancer diagnosis, prognosis and related mechanism research. And it will also provide important references for individual diagnoses and treatments of cancer.

RNA可变剪接是调节基因表达和产生蛋白质多样性的重要机制，与癌症等重大疾病的发生发展密切相关。异常RNA剪接产生的变体有望作为癌症诊断或预后监测的潜在标志物。本项目围绕RNA可变剪接高灵敏高特异性测定及原位荧光成像方法构建开展研究，探索剪接变体在细胞中的分布表达差异及其与癌症相关性。开展核酸纳米结构构筑研究，利用核酸纳米结构作为信号放大元件，结合RNA可变剪接引起的下游分子结构特点，研究RNA可变剪接变体特异性分子识别信号转换及信号放大机制，建立RNA可变剪接事件高灵敏高选择性测定以及细胞内单个变体分子原位荧光成像新方法。进而系统研究肺癌细胞内RNA可变剪接变体的表达与分布差异，分析它们与肺癌相关性。该项目研究将为癌症等重大疾病的诊断、预后监测及发病机理研究提供重要的理论与技术支持，为癌症个体化诊疗提供重要参考。

RNA可变剪接是调节基因表达和产生蛋白质多样性的重要机制，和癌症、遗传性疾病等重大疾病的发生发展密切相关。发展高灵敏度、高选择性、可以多组分分析的RNA可变剪接变体原位测定方法，在可变剪接与疾病相互关系研究以及正确理解基因表达过程、探索疾病发病机理和进行疾病诊断中具有重要意义。本项目围绕构建高灵敏高特异性的RNA可变剪接测定及原位荧光成像方法，研究了多种功能材料和核酸纳米结构的构筑及其光学、催化、生物相容等相关性能，探索了核酸纳米结构介导的分子识别信号转换与放大方法；并进一步结合RNA及可变剪接变体分子特点，以核酸纳米结构为信号转换与放大元件，建立了系列RNA测定及成像新方法，开展了RNA及可变剪接变体的原位荧光成像分析，并探索了细胞内不同剪接变体的多组分测定及其表达情况。本项目为RNA可变剪接相关的生物医学研究提供了重要的技术及理论参考。通过本项目研究，已经发表了12篇学术论文，培养或协助培养了1名博士研究生和10名硕士研究生。