Construction of synthetic circuits that can artificially establish endogenous gene connections is essential to introduce new phenotypes for cellular behaviors. Given the diversity of endogenous genes, it lacks a general and easy-to-design toolbox to manipulate the genetic network. Here we present a type of self-assembly-induced RNA circuit that can directly build regulatory connections between endogenous genes. Inspired from the natural assembling process of guide RNA in the CRISPR/Cas9 complex, this design employs an independent trigger RNA strand to induce the formation of a ternary guide RNA assembly for functional control of CRISPR/Cas9. With this general principle, expressional regulations of endogenous genes can be controlled by totally independent endogenous small RNAs and mRNAs in E. coli via activatable CRISPR/Cas9 function. Moreover, the cellular phenotype of E. coli is successfully programmed with introduction of new gene connections. In addition, the functionality of this design is also verified in the mammalian system. This self-assembly-based RNA circuit exhibits a great flexibility and simplicity of design and provides a unique approach to build endogenous gene connections, which paves a broad way toward manipulation of cellular genetic networks.

中文翻译：

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通过 RNA 自组装控制的 CRISPR/Cas9 功能建立内源基因连接

构建可以人工建立内源基因连接的合成电路对于为细胞行为引入新的表型至关重要。鉴于内源基因的多样性，它缺乏一个通用且易于设计的工具箱来操纵遗传网络。在这里，我们提出了一种自组装诱导的 RNA 回路，可以直接在内源基因之间建立调节连接。受 CRISPR/Cas9 复合物中引导 RNA 的自然组装过程的启发，该设计采用独立的触发 RNA 链来诱导形成三元引导 RNA 组装，以实现对 CRISPR/Cas9 的功能控制。根据这一一般原则，内源基因的表达调控可以由大肠杆菌中完全独立的内源小 RNA 和 mRNA 控制。通过可激活的 CRISPR/Cas9 功能。此外，通过引入新的基因连接，成功地对大肠杆菌的细胞表型进行了编程。此外，该设计的功能性也在哺乳动物系统中得到验证。这种基于自组装的 RNA 电路表现出极大的灵活性和设计的简单性，并提供了一种独特的方法来建立内源基因连接，这为操纵细胞遗传网络铺平了道路。