Design and Characterization of a Micro RNA-200c Detecting Broccoli Fluorescent Light-up Aptamer

In the last decade the important role of small non-coding RNAs such as micro RNAs (miRs) in gene regulation in healthy and disease states became more and more evident. The miR-200-family of miRs has been shown to play a critical role in many diseases such as cancer and neurodegenerative disorders and could be potentially important for diagnosis and treatment. However, the size of miRs of about ~21-23 nt provide challenges for their investigation. Here we report the conversion and optimization of the Broccoli fluorescent light-up RNA-aptamer into a specific sensor for miR-200c using a strand-displacement design principle. This aptamer can differentiate miR-200c from its family members whose sequence differ by more than one nucleotide. By adding this in vitro transcribed aptamer to RNA extracts from human cells, we can detect miR-200c in vitro in a plate reader assay.