Studying RNA Using Single Molecule Fluorescence Resonance Energy Transfer

A modified guanosine phosphoramidite for click functionalization of RNA on the sugar edge

A propargyl containing guanosine phosphoramidite was synthesized and incorporated into siRNA, enabling click-ligation with an azido fluorophore onto the nucleobase sugar edge. Duplex stability was not affected by labeling at this new site, which allowed deconvolution of the effects of label, structure and attachment site on RNAi activity.