Department of Pharmacology, Weill Cornell Medicine, Cornell University, New York, New York 10065, United States.
Beijing Institutes of Life Science, Chinese Academy of Sciences, Beijing 100101, P. R. China.
J Am Chem Soc. 2022 Mar 30;144(12):5471-5477. doi: 10.1021/jacs.1c13583. Epub 2022 Mar 16.
DNA and RNA can spontaneously self-assemble into various structures, including aggregates, complexes, and ordered structures. The self-assembly reactions cannot be genetically encoded to occur in living mammalian cells since the double-stranded nucleic acids generated by current self-assembly approaches are unstable and activate innate RNA immunity pathways. Here, we show that recently described dimeric aptamers can be used to create RNAs that self-assemble and create RNA and RNA-protein assemblies in cells. We find that incorporation of five copies of Corn, a dimeric fluorogenic RNA aptamer, into an RNA causes the RNA to form large clusters in cells, reflecting multivalent RNA-RNA interactions enabled by these RNAs. Here, we also describe a second dimeric fluorogenic aptamer, Beetroot, which shows partial sequence similarity to Corn. Both Corn and Beetroot form homodimers with themselves but do not form Corn-Beetroot heterodimers. We thus use Corn and Beetroot to encode distinct RNA-protein assemblies in the same cells. Overall, these studies provide an approach for inducing RNA self-assembly, enable multiplexing of distinct RNA assemblies in cells, and demonstrate that proteins can be recruited to RNA assemblies to genetically encode intracellular RNA-protein assemblies.
DNA 和 RNA 可以自发地自组装成各种结构，包括聚集体、复合物和有序结构。由于当前自组装方法产生的双链核酸不稳定，并激活先天 RNA 免疫途径，因此这些自组装反应不能通过遗传编码在活的哺乳动物细胞中发生。在这里，我们表明，最近描述的二聚体适体可用于创建可在细胞中自组装并形成 RNA 和 RNA-蛋白质组装的 RNA。我们发现，将 5 个拷贝的 Corn（一种二聚体荧光 RNA 适体）掺入到 RNA 中会导致 RNA 在细胞中形成大的簇，这反映了这些 RNA 所实现的多价 RNA-RNA 相互作用。在这里，我们还描述了第二个二聚体荧光适体 Beetroot，它与 Corn 具有部分序列相似性。Corn 和 Beetroot 都可以与自身形成同源二聚体，但不能形成 Corn-Beetroot 异源二聚体。因此，我们使用 Corn 和 Beetroot 在相同的细胞中编码不同的 RNA-蛋白质组装体。总体而言，这些研究提供了一种诱导 RNA 自组装的方法，能够在细胞中对不同的 RNA 组装体进行多重化，并证明蛋白质可以被招募到 RNA 组装体中，从而对细胞内的 RNA-蛋白质组装体进行遗传编码。
