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Kwon, Hyokyoung,Kim, Minjeong,Seo, Yunmi,Moon, Yae Seul,Lee, Hwa Jeong,Lee, Kyuri,Lee, Hyukjin Elsevier 2018 Biomaterials Vol.156 No.-
<P><B>Abstract</B></P> <P>The field of gene therapy has evolved over the past two decades after the first introduction of nucleic acid drugs, such as plasmid DNA (pDNA). With the development of <I>in vitro</I> transcription (IVT) methods, synthetic mRNA has become an emerging class of gene therapy. IVT mRNA has several advantages over conventional pDNA for the expression of target proteins. mRNA does not require nuclear localization to mediate protein translation. The intracellular process for protein expression is much simpler and there is no potential risk of insertion mutagenesis. Having these advantages, the level of protein expression is far enhanced as comparable to that of viral expression systems. This makes IVT mRNA a powerful alternative gene expression system for various applications in regenerative medicine. In this review, we highlight the synthesis and preparation of IVT mRNA and its therapeutic applications. The article includes the design and preparation of IVT mRNA, chemical modification of IVT mRNA, and therapeutic applications of IVT mRNA in cellular reprogramming, stem cell engineering, and protein replacement therapy. Finally, future perspectives and challenges of IVT mRNA are discussed.</P>
Jang, Bora,Kim, Boyoung,Kim, Hyunsook,Kwon, Hyokyoung,Kim, Minjeong,Seo, Yunmi,Colas, Marion,Jeong, Hansaem,Jeong, Eun Hye,Lee, Kyuri,Lee, Hyukjin American Chemical Society 2018 NANO LETTERS Vol.18 No.7
<P>Enzymatic synthesis of RNA nanostructures is achieved by isothermal rolling circle transcription (RCT). Each arm of RNA nanostructures provides a functional role of Dicer substrate RNA inducing sequence specific RNA interference (RNAi). Three different RNAi sequences (GFP, RFP, and BFP) are incorporated within the three-arm junction RNA nanostructures (Y-RNA). The template and helper DNA strands are designed for the large-scale in vitro synthesis of RNA strands to prepare self-assembled Y-RNA. Interestingly, Dicer processing of Y-RNA is highly influenced by its physical structure and different gene silencing activity is achieved depending on its arm length and overhang. In addition, enzymatic synthesis allows the preparation of various Y-RNA structures using a single DNA template offering on demand regulation of multiple target genes.</P> [FIG OMISSION]</BR>