Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells.
Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells.
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DOI：
10.1093/nar/gkx298
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发表时间：
2017-07-07
影响因子：
14.9
通讯作者：
Saito H
中科院分区：
文献类型：
--
作者：
Kawasaki S;Fujita Y;Nagaike T;Tomita K;Saito H
合成生物学在未来的治疗应用中具有巨大的潜力，包括通过检测蛋白质信号和产生所需输出来进行自主细胞编程。合成RNA设备有望实现这一目的。然而，由于难以检测细胞内的内源性蛋白质，可用装置的数量有限。在这里，我们展示了一种构建合成mRNA装置的策略，该装置可以检测活细胞中的内源性蛋白质，控制翻译并区分细胞类型。我们设计了蛋白质结合适体，其活性构象的二级结构具有增加的稳定性。设计的装置可以有效地响应包括人类LIN28A和U1A蛋白在内的靶蛋白，而原始适体未能做到这一点。此外，将LIN28A响应装置的mRNA递送到人诱导多能干细胞（hiPSC）中揭示了我们可以通过定量内源性LIN28A蛋白表达水平来区分活的hiPSC和分化的细胞。因此，我们的内源性蛋白质驱动的RNA设备确定活细胞状态，并根据细胞内蛋白质信息对哺乳动物细胞进行编程。
Synthetic biology has great potential for future therapeutic applications including autonomous cell programming through the detection of protein signals and the production of desired outputs. Synthetic RNA devices are promising for this purpose. However, the number of available devices is limited due to the difficulty in the detection of endogenous proteins within a cell. Here, we show a strategy to construct synthetic mRNA devices that detect endogenous proteins in living cells, control translation and distinguish cell types. We engineered protein-binding aptamers that have increased stability in the secondary structures of their active conformation. The designed devices can efficiently respond to target proteins including human LIN28A and U1A proteins, while the original aptamers failed to do so. Moreover, mRNA delivery of an LIN28A-responsive device into human induced pluripotent stem cells (hiPSCs) revealed that we can distinguish living hiPSCs and differentiated cells by quantifying endogenous LIN28A protein expression level. Thus, our endogenous protein-driven RNA devices determine live-cell states and program mammalian cells based on intracellular protein information.