新型高通量microRNA免标记芯片技术研究

microRNA是近年来发现的一种长度约22个碱基的短链RNA，在动植物和病毒中广泛存在，对mRNA 的表达进行调控，与包括发育、分化、凋亡、脂肪代谢、病毒感染和癌症等多种重要生物学过程有密切的联系，是近十年来的一个研究热点。microRNA 芯片作为一种高通量的microRNA 表达谱检测工具被广泛使用。但是，microRNA 的长度太短，标记比较困难，需要相当多的操作步骤来实现，增加了成本和人工操作时间。针对上述问题，我们希望发展一种新的microRNA 非标记高通量芯片检测技术，可以对microRNA 进行直接检测。该方法是基于碱基堆积杂交的原理，加入一种带有荧光的通用标签，采用杂交触发荧光的策略，在杂交发生的同时产生荧信号。 研究内容包括芯片设计和制作、性能评价和动植物细胞及组织的microRNA 表达分析等。

microRNAs(miRNAs)are a class of small single-stranded noncoding RNAs, about 22 nucleotides long that have been found in animals, plants, viruses and other species. miRNAs have important roles in plant and animal development, apoptosis, growth control and hematopoietic differentiation. Owing to the impressive progress in understanding the correlations between miRNA expression and diseases, miRNAs have been considered novel biomarkers of various human diseases including cancer, viral diseases, cardiovascular disorders, metabolic disturbances, etc. The microarray is a powerful platform for high-throughput miRNAs gene expression analysis. While the short length of miRNA present major challenges to sample labeling. Several strategies have been developed to overcome this difficulty. Most of them require hours of manual steps to label the miRNA, which are laborious, prone to error and usually expensive. We have designed a hybridization-triggered fluorescence strategy for label-free high-throughput miRNA expression profiling, which is superior to most existing chemical and enzymatic labeling methods.