Fluorescence Activation Imaging of Cytochrome c Released from Mitochondria Using Aptameric Nanosensor
Fluorescence Activation Imaging of Cytochrome c Released from Mitochondria Using Aptameric Nanosensor
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DOI：
10.1021/ja511988w
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发表时间：
2015-01-21
影响因子：
15
通讯作者：
Li, Yingfu
中科院分区：
文献类型：
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作者：
Chen, Ting-Ting;Tian, Xue;Li, Yingfu
我们开发了一种用于细胞色素c （Cyt c）荧光激活成像的适体纳米传感器。荧光成像工具能够可视化凋亡信号传导中的关键分子，对于细胞生物学和临床治疗是必不可少的。Cyt c是细胞凋亡的主要介质。然而，荧光成像工具允许直接可视化Cyt c在活细胞中的易位目前尚未实现。我们首次报道了一种纳米传感器工具的实现，该工具能够对细胞凋亡过程中线粒体释放的Cyt c进行直接荧光激活成像。该策略依赖于在聚乙二醇化石墨烯纳米片上组装荧光团标记的DNA适体而构建的纳米传感器的空间选择性细胞质递送。胞质释放的Cyt c能够将适体与石墨烯分离，并触发激活的荧光信号。该纳米传感器具有高灵敏度和选择性、快速响应、大信号背景比的体外细胞内检测Cyt - c的特性，还可以实时可视化Cyt - c释放动力学和直接识别细胞凋亡调节因子。所开发的纳米传感器可以为细胞凋亡研究提供一个非常有价值的工具，并催化细胞凋亡介导生物学的基本问题。
We have developed an aptameric nanosensor for fluorescence activation imaging of cytochrome c (Cyt c). Fluorescence imaging tools that enable visualization of key molecular players in apoptotic signaling are essential for cell biology and clinical theranostics. Cyt c is a major mediator in cell apoptosis. However, fluorescence imaging tools allowing direct visualization of Cyt c translocation in living cells have currently not been realized. We report for the first time the realization of a nanosensor tool that enables direct fluorescence activation imaging of Cyt c released from mitochondria in cell apoptosis. This strategy relies on spatially selective cytosolic delivery of a nanosensor constructed by assembly of a fluorophore-tagged DNA aptamer on PEGylated graphene nanosheets. The cytosolic release of Cyt c is able to dissociate the aptamer from graphene and trigger an activated fluorescence signal. The nanosensor is shown to exhibit high sensitivity and selectivity, rapid response, large signal-to-background ratio for in vitro, and intracellular detection of Cyt c. It also enables real-time visualization of the Cyt c release kinetics and direct identification of the regulators for apoptosis. The developed nanosensor may provide a very valuable tool for apoptotic studies and catalyze the fundamental interrogations of Cyt c-mediated biology.