Fluorescene Resonance Energy Transfer (FRET) in RNA Binding Studies: In-vitro Studies on Aptamer ? Induced FRET Between ?N22 Tagged Fluorescent Protein Variants

This work tries to show an attempt in the ribonucleic acid (RNA) tagging field. With many different types of RNAs found in a living cell, interest has developed in not only to study their function but also their precise location and time of action in a cellular process. Tagging and monitoring an RNA in a live cell using a fluorescent marker is a challenging field and one that would open the doors to many unanswered questions on RNA monitoring, conformational change and others. Fluorescent proteins are widely used for tagging and visualizing biomolecules in vivo or in solution. With the use of fluorescent proteins that are good FRET (Fluorescence Resonance Energy Transfer) partners, this work has tried to study if a FRET ratio change occurs when the boxB RNA folds in solution on binding of the boxB RNA binding peptides, ?N22. This work can be helpful to other researchers trying to address similar questions and can be useful in knowing what and also what not to do in their experiments.