Versatile strategy using vaccinia virus-capping enzyme to synthesize functional 5' cap-modified mRNAs.

The potential of synthetic mRNA as a genetic carrier has increased its application in scientific fields. Because the 5' cap regulates the stability and translational activity of mRNAs, there are concerted efforts to search for and synthesize chemically-modified 5' caps that improve the functionality of mRNA. Here, we report an easy and efficient method to synthesize functional mRNAs by modifying multiple 5' cap analogs using a vaccinia virus-capping enzyme. We show that this enzyme can introduce a variety of GTP analogs to the 5' end of RNA to generate 5' cap-modified mRNAs that exhibit different translation levels. Notably, some of these modified mRNAs improve translation efficiency and can be conjugated to chemical structures, further increasing their functionality. Our versatile method to generate 5' cap-modified mRNAs will provide useful tools for RNA therapeutics and biological research.

Synthetic mRNA-Based Systems in Mammalian Cells.

Living organisms are programmed to perform multiple functions by sensing intra- and extra-cellular environments and by controlling gene expressions. Synthetic biologists aim to program cells by mimicking, designing, and constructing genetic circuits. Synthetic mRNA-based genetic switches and circuits have attracted attention for future therapeutic applications because of their safety and functional diversity. Here, the mRNA-based switches and circuits that detect specific microRNAs or proteins expressed in a target cell to control transgene expression and cell fate are reviewed. Future perspectives of artificial RNA systems for cell engineering will also be addressed.

RNA nanostructures and scaffolds for biotechnology applications.

RNA plays important roles in the regulation of gene expressions and other cellular functions. It functions as both as an informational carrier and a nanomachine due to its complementary base-pairing ability and complexed three-dimensional structure. Several nanostructures have been designed and constructed by exploiting these natural RNA properties. In this review, we will introduce the design principles of RNA nanostructures and their biotechnology applications as molecular scaffolds. RNA-based molecular scaffolds can control the accumulation and interaction of target proteins at nanometer-scale to regulate the function of bacterial and mammalian cells. Combining useful property of RNA as a nano-material and a molecular scaffold may provide us powerful tools in biological research, bioengineering, and future medicine.