Light-controllable RNA-protein devices for translational regulation of synthetic mRNAs in mammalian cells
Light-controllable RNA-protein devices for translational regulation of synthetic mRNAs in mammalian cells
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用于哺乳动物细胞中合成 mRNA 翻译调节的光控 RNA 蛋白装置
DOI：
10.1016/j.chembiol.2021.01.002
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发表时间：
2021
影响因子：
8.6
通讯作者：
Hirohide Saito
中科院分区：
文献类型：
--
作者：
Hideyuki Nakanishi;Tatsuyuki Yoshii;Shunsuke Kawasaki;Karin Hayashi;Keita Tsutsui;Choji Oki;Shinya Tsukiji;Hirohide Saito
转基因表达的光调控因其高的时空分辨率而成为哺乳动物合成生物学研究的有效途径之一。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，modRNA递送的转基因的光遗传学控制比DNA递送的转基因的光遗传学控制困难得多。在这里，我们开发了两种类型的与modRNA兼容的光控翻译激活系统。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异二聚化。另一个是由一个不稳定的结构域融合的翻译激活蛋白和光笼稳定剂。不稳定型不仅可用于modRNA的翻译激活，而且可用于modRNA的翻译抑制。这些光控翻译系统将拓展哺乳动物合成生物学研究的应用。
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.