Department of Molecular Biology, Cell Biology and Biochemistry, Brown University, Providence, Rhode Island, USA.
Nat Methods. 2011 Dec 25;9(2):189-94. doi: 10.1038/nmeth.1827.
Informational recoding by adenosine-to-inosine RNA editing diversifies neuronal proteomes by chemically modifying structured mRNAs. However, techniques for analyzing editing activity on substrates in defined neurons in vivo are lacking. Guided by comparative genomics, here we reverse-engineered a fluorescent reporter sensitive to Drosophila melanogaster adenosine deaminase that acts on RNA (dADAR) activity and alterations in dADAR autoregulation. Using this artificial dADAR substrate, we visualized variable patterns of RNA-editing activity in the Drosophila nervous system between individuals. Our results demonstrate the feasibility of structurally mimicking ADAR substrates as a method to regulate protein expression and, potentially, therapeutically repair mutant mRNAs. Our data suggest variable RNA editing as a credible molecular mechanism for mediating individual-to-individual variation in neuronal physiology and behavior.
腺苷到肌苷的 RNA 编辑通过化学修饰结构 mRNA 来改变神经元的蛋白质组。然而，目前缺乏用于分析体内特定神经元中底物编辑活性的技术。在比较基因组学的指导下，我们在这里反向设计了一个对果蝇腺苷脱氨酶起作用的 RNA (dADAR) 活性和 dADAR 自我调节变化敏感的荧光报告基因。利用这种人工 dADAR 底物，我们在果蝇神经系统中个体之间可视化了 RNA 编辑活性的可变模式。我们的结果证明了结构模拟 ADAR 底物作为调节蛋白质表达的一种方法的可行性，并且可能具有治疗修复突变 mRNA 的潜力。我们的数据表明，可变 RNA 编辑是介导神经元生理和行为个体间差异的可信分子机制。
