Department of Life Science Frontiers, Center for iPS Cell Research and Application, Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan; Department of Biofunction Research, Institute of Biomaterials and Bioengineering, Tokyo Medical and Dental University (TMDU), 2-3-10 Kanda-Surugadai, Chiyoda-ku, Tokyo 101-0062, Japan.
Department of Life Science and Applied Chemistry, Nagoya Institute of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan; PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
Cell Chem Biol. 2021 May 20;28(5):662-674.e5. doi: 10.1016/j.chembiol.2021.01.002. Epub 2021 Jan 27.
The photo-regulation of transgene expression is one effective approach in mammalian synthetic biology due to its high spatial and temporal resolution. While DNAs are mainly used as vectors, modified RNAs (modRNAs) are also useful for medical applications of synthetic biology, because they can avoid insertional mutagenesis and immunogenicity. However, the optogenetic control of modRNA-delivered transgenes is much more difficult than that of DNA-delivered transgenes. Here, we develop two types of photo-controllable translational activation systems that are compatible with modRNAs. One is composed of a heterodimerization domain-fused split translational activator protein and a photocaged heterodimerizer. The other is composed of a destabilizing domain-fused translational activator protein and a photocaged stabilizer. The destabilized type can be used for not only translational activation but also translational repression of the modRNAs. These photo-controllable translation systems will expand the application of mammalian synthetic biology research.
光调控转基因表达是哺乳动物合成生物学中一种有效的方法，因为它具有高时空分辨率。虽然 DNA 主要用作载体，但修饰的 RNA（modRNA）也可用于合成生物学的医学应用，因为它们可以避免插入突变和免疫原性。然而，与 DNA 递送的转基因相比，光遗传学控制 modRNA 递送的转基因要困难得多。在这里，我们开发了两种与 modRNA 兼容的光可控翻译激活系统。一种是由异二聚化结构域融合的分裂翻译激活蛋白和光笼合异二聚体组成。另一种是由去稳定化结构域融合的翻译激活蛋白和光笼合稳定剂组成。去稳定化类型不仅可用于 modRNA 的翻译激活，还可用于翻译抑制。这些光可控翻译系统将扩展哺乳动物合成生物学研究的应用。
