Fluorogenic RNA aptamers: Exploring applications for nucleic acid detection

Worldwide concern for disease detection has led to demand for sensitive, specific, and simple pathogen detection methods. In recent years, methods for detecting nucleic acids, including molecular beacon reporters and the isothermal amplification of nucleic acids have become popular. Highly fluorescent RNA Mango aptamers have been developed that can operate in salt concentrations required for in vitro systems. Presented here are two novel applications of RNA Mango aptamers for the detection of nucleic acids. First, by engineering RNA Mango, a molecular beacon like switch was developed that maintains minimal fluorescence in the absence of RNA target, and that dramatically increases in fluorescence upon target addition. Second, by incorporating Mango aptamers into an existing isothermal amplification method (NASBA), attoMolar concentrations of RNA can be detected with high sensitivity and specificity. These Mango tools improve upon current RNA detection technologies and provide simple fluorescent methods for detecting RNA.