| 초록 |
Plasmid DNA (pDNA) is often used as a genetic material to induce target protein expression in mammalian cells. Recently, messenger RNA (mRNA) has drawn much attention as an alternative genetic material that can be expressed in mammalian cells instead of pDNA. mRNA is a genetic material synthesized by transcription of DNA template and the resultant single strand RNA molecules are used to induce the protein expression in mammalian cells. As compared to the conventional pDNA delivery, mRNA delivery has multiple advantages such as 1) no nuclear localization is required, 2) less genetic stress on the host cells, and 3) fast on-set target gene expression. However, due to its nature, unstable single strand RNA structure is vulnerable to enzyme degradation and immune system. In this study, we have prepared mRNA by in vitro transcription (IVT) methods along with linearization of pDNA. In addition, chemical modification of mRNA has been attempted by utilizing modified nucleoside, 5’ capping, and 3’ poly (A) tail to avoid immune system and degradation. Transfection and expression of IVT mRNA have been confirmed in mouse embryonic fibroblast (MEF) cells.
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