Anal Chem. 2011 Sep 15;83(18):6935-9. doi: 10.1021/ac201903n. Epub 2011 Aug 26.
MicroRNAs (miRNAs) are regulatory small RNAs that have important roles in numerous developmental, metabolic, and disease processes of plants and animals. The individual levels of miRNAs can be useful biomarkers for cellular events or disease diagnosis. Thus, innovative new tools for rapid, specific, and sensitive detection of miRNAs are an important field of research. Using the fluorescence properties of DNA-nanosilver clusters (DNA/AgNC), we have designed a DNA/AgNC probe that can detect the presence of target miRNA. Here, we show that the red fluorescence of the DNA/AgNC probe is diminished upon the presence of target miRNA without pre- or postmodification, addition of extra enhancer molecules, and labeling. The DNA/AgNC probe emission was lowest when the complementary miRNA target was present and was significantly higher for four other control miRNA sequences. Also, when adding whole plant endogenous RNA to the DNA/AgNC probe, the emission was significantly higher for the mutant where miRNA was deficient. On the basis of these findings, we suggest that these DNA/AgNC probes could be developed into a new, simple, inexpensive, and instant technique for miRNAs detection.
微小 RNA(miRNAs)是调节性小 RNA,在动植物的许多发育、代谢和疾病过程中发挥重要作用。miRNAs 的个体水平可以作为细胞事件或疾病诊断的有用生物标志物。因此,用于快速、特异和灵敏检测 miRNAs 的创新新工具是一个重要的研究领域。利用 DNA-纳米银簇(DNA/AgNC)的荧光性质,我们设计了一种可以检测靶 miRNA 存在的 DNA/AgNC 探针。在这里,我们表明,在没有预或后修饰、添加额外增强子分子和标记的情况下,存在靶 miRNA 会使 DNA/AgNC 探针的红色荧光减弱。当互补的 miRNA 靶标存在时,DNA/AgNC 探针的发射最低,而对于其他四个对照 miRNA 序列则显著升高。此外,当向 DNA/AgNC 探针中添加植物内源 RNA 时,miRNA 缺失的突变体的发射显著升高。基于这些发现,我们认为这些 DNA/AgNC 探针可以开发成一种新的、简单的、廉价的、即时的 miRNA 检测技术。