Establishing artificial gene connections through RNA displacement-assembly-controlled CRISPR/Cas9 function.
Establishing artificial gene connections through RNA displacement-assembly-controlled CRISPR/Cas9 function.
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DOI：
10.1093/nar/gkad558
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发表时间：
2023-08-11
影响因子：
14.9
通讯作者：
Xu, Liang
中科院分区：
文献类型：
--
作者：
Wang, Wei-Jia;Lin, Jiao;Wu, Chao-Qun;Luo, Ai-Ling;Xing, Xiwen;Xu, Liang
构建可以重新编程遗传网络和信号通路的合成电路是操纵生物系统的长期目标。然而，由于内源性RNA的序列独立性和结构多样性，在它们之间建立人工遗传通讯仍然具有很大的挑战性。在这里，我们报告了一个基于RNA的合成电路，可以建立在土方ichiacoli和哺乳动物细胞的内源性基因的表达之间的调控联系。该设计采用置换-组装方法来调节指导RNA的活性以用于CRISPR/Cas9的功能控制。我们的实验证明了这种RNA回路在建立最初不相关基因表达之间的人工连接方面的巨大有效性。外源和天然存在的RNA，包括小/微RNA和长mRNA，都能够通过这种方法控制另一种内源基因的表达。此外，通过我们设计的合成电路，还成功地在哺乳动物细胞内建立了一条人工信号通路来控制细胞凋亡。本研究为构建人工合成的RNA回路提供了一种通用策略，该策略可以将人工连接引入哺乳动物细胞的遗传网络并改变细胞表型。
Construction of synthetic circuits that can reprogram genetic networks and signal pathways is a long-term goal for manipulation of biosystems. However, it is still highly challenging to build artificial genetic communications among endogenous RNA species due to their sequence independence and structural diversities. Here we report an RNA-based synthetic circuit that can establish regulatory linkages between expression of endogenous genes in both Escherichiacoli and mammalian cells. This design employs a displacement–assembly approach to modulate the activity of guide RNA for function control of CRISPR/Cas9. Our experiments demonstrate the great effectiveness of this RNA circuit for building artificial connections between expression of originally unrelated genes. Both exogenous and naturally occurring RNAs, including small/microRNAs and long mRNAs, are capable of controlling expression of another endogenous gene through this approach. Moreover, an artificial signal pathway inside mammalian cells is also successfully established to control cell apoptosis through our designed synthetic circuit. This study provides a general strategy for constructing synthetic RNA circuits, which can introduce artificial connections into the genetic networks of mammalian cells and alter the cellular phenotypes.
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影响因子：
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通讯作者：
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