Computing in mammalian cells with nucleic acid strand exchange.
Computing in mammalian cells with nucleic acid strand exchange.
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DOI：
10.1038/nnano.2015.278
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发表时间：
2016-03
影响因子：
38.3
通讯作者：
Seelig G
中科院分区：
文献类型：
--
作者：
Groves B;Chen YJ;Zurla C;Pochekailov S;Kirschman JL;Santangelo PJ;Seelig G
DNA链置换已被广泛应用于无细胞环境下的分子电路、电机和传感器的设计。最近，有研究表明，这项技术也可以在生物环境中运行，但能力仍然有限。在这里，我们希望使链置换和交换反应适应哺乳动物细胞，并报告可以直接与天然mRNA相互作用的DNA电路。我们首先基于四向链交换反应优化荧光报告的细胞性能，并通过系统地改变分子结构、化学和传递方法来确定可靠的设计原则。接下来，我们开发并测试了基于四路串交换的AND和OR逻辑门，论证了多输入逻辑的可行性。最后，我们证实了功能性的siRNA可以通过链交换被激活，并使用天然的mRNA作为可编程的支架来共定位GATES并以亚细胞分辨率可视化它们的操作。
DNA strand displacement has been widely used for the design of molecular circuits, motors, and sensors in cell-free settings. Recently, it has been shown that this technology can also operate in biological environments, but capabilities remain limited. Here, we look to adapt strand displacement and exchange reactions to mammalian cells and report DNA circuitry that can directly interact with a native mRNA. We began by optimizing the cellular performance of fluorescent reporters based on four-way strand exchange reactions and identified robust design principles by systematically varying the molecular structure, chemistry and delivery method. Next, we developed and tested AND and OR logic gates based on four-way strand exchange, demonstrating the feasibility of multi-input logic. Finally, we established that functional siRNA could be activated through strand exchange, and used native mRNA as programmable scaffolds for co-localizing gates and visualizing their operation with subcellular resolution.