Graphene oxide size and oxidation degree govern its supramolecular interactions with siRNA.

Several studies have demonstrated the ability of graphene oxide (GO) to efficiently adsorb small-interfering RNA (siRNA) on its surface and to transport it into cells. However, studies on whether and how siRNA interacts with GO are still inconclusive. In this context, understanding the interaction between GO and siRNA is fundamental to design new efficient gene silencing tools. In this work, the interactions between GO and siRNA molecules were systematically investigated. We focused on how the GO size, oxygenated groups present on the surface and chemical functionalization affect the double helix siRNA structure, using gel electrophoresis, UV-Vis spectroscopy, fluorescence resonance energy transfer (FRET) and circular dichroism (CD). We found that the siRNA secondary structure was clearly altered by the interaction with GO flakes. In addition, we were able to correlate the double strand damage with the size and the oxygenated groups present on the GO sheets. Finally, we demonstrated that GO functionalized with low molecular weight polyethyleneimine (PEI, 800 Da) is able to protect siRNA from structural modifications. We believed that this research effort will improve our understanding of the behavior of GO/siRNA complexes, and thus facilitate the design of appropriate bio/nanointerfaces and new efficient gene silencing systems.