Abstract
After the discovery of small interference RNA (siRNA), nanostructured siRNA delivery systems have been introduced to achieve an efficient regulation of the target gene expression. Here we report a new siRNA-generating two dimensional nanostructure in a formation of nanosized sheet. Inspired by tunable mechanical and functional properties of the previously reported RNA membrane, siRNA nanosized sheets (siRNA-NS) with multiple Dicer cleavage sites were prepared. The siRNA-NS has two dimensional structure, providing a large surface area for Dicer to cleave the siRNA-NS for the generation of functional siRNAs. Furthermore, downregulation of the cellular target gene expression was achieved by delivery of siRNA-NS without chemical modification of RNA strands or conjugation to other substances.
Highlights
After the discovery of small interference ribonucleic acid (RNA), nanostructured small interference RNA (siRNA) delivery systems have been introduced to achieve an efficient regulation of the target gene expression
RNA has been recently used as a generic material for nano- and micro-architectures in the scope of materials engineering to utilize a wide range of its functions[2,3,4]
A novel approach to prepare RNA membrane without polymer supports or complexation was recently introduced[14]. This was the first example of a macroscopic RNA membrane, which is composed of RNA strands and has tunable mechanical properties and biological functions
Summary
To synthesize siRNA-NS for mediating RNAi, two types of circular DNAs with rationally designed sequences were first generated using the method reported elsewhere[14,15]. RNA membrane was torn up into the nanosized sheets (Fig. 1b). After a brief ultrasonication for 20 s, RNA membrane become fractured into large pieces still visible by naked eyes when stained with GelRed. After further sonication, dominating products were microsized (Fig. 2b). The high magnification SEM image of the split site of the RNA membrane in the process of production of siRNA-NS reveals that the RNA membrane is composed of multiple thinner layers. Further analysis with energy-dispersive X-ray spectroscopy (EDX) revealed siRNA-NS is composed of carbon (C), nitrogen (N), oxygen (O) and phosphorous (P), indicating siRNA-NS is made of RNA strands (Fig. 2f)
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