All-in-One Synchronized DNA Nanodevices Facilitating Multiplexed Cell Imaging
All-in-One Synchronized DNA Nanodevices Facilitating Multiplexed Cell Imaging
复制标题
多合一同步 DNA 纳米器件促进多重细胞成像
DOI：
10.1021/acs.analchem.9b00089
复制
发表时间：
2019-04-02
影响因子：
7.4
通讯作者：
Zhao，Yongxi
中科院分区：
文献类型：
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作者：
Xue，Jing;Chen，Feng;Zhao，Yongxi
多功能 DNA 纳米器件可以执行越来越多的任务，其应用范围从体外生物标志物检测到原位细胞成像。然而，大多数开发的模块由一系列分裂的构建块组成，这些构建块在复杂的细胞微环境（胞吞途径、扩散限制的细胞质等）中遭受异步行为，导致化学计量信息的丢失和额外的后组装过程。在此，我们构建了一体化 DNA 纳米器件来实现同步多重成像。所有 DNA 组件，包括两组探针模块（每组都包含特定目标的步行器，即具有化学损坏碱基的发夹轨道），均在单个金纳米粒子上进行修饰。这种设计不仅使它们能够整合内化到细胞中，避免不同构建模块的不均匀分布并增加相互作用模块的局部浓度，而且还避免了粘性细胞质中随机扩散的影响。在特异性识别细胞内靶标（例如 RNA 和蛋白质）后，一些细胞内酶原位驱动模块的同步运动（所有模块均在颗粒上），从而促进同步、多重细胞成像。最后，所提出的一体化纳米装置成功应用于监测细胞内 microRNA-21 和端粒酶的表达水平。灵活的设计可以扩展到检测其他细胞质分子并通过简单地改变序列来监测相关途径。
Multifunctional DNA nanodevices perform ever more tasks with applications ranging from in vitro biomarker detection to in situ cell imaging. However, most developed ones consist of a series of split building blocks, which suffer from asynchronous behaviors in complicated cellular microenvironments (endocytosis pathway, diffusion-limited cytoplasm, etc.), causing the loss of stoichiometric information and additional postassembly processes. Herein, we constructed all-in-one DNA nanodevices to achieve synchronous multiplexed imaging. All DNA components, including two sets of probe modules (each containing target-specific walkers, i.e., hairpin tracks with chemically damaged bases), are modified on individual gold nanoparticles. This design not only enables their integrated internalization into cells, circumventing inhomogeneous distribution of different building blocks and increasing the local concentrations of the interacting modules, but also avoids the impact of stochastic diffusion in viscous cytoplasm. A couple of intracellular enzymes in situ actuate the synchronized motion of the modules, all on-particle, after specific recognition of intracellular targets (such as RNAs and proteins), thus facilitating synchronized, multiplexed cell imaging. Finally, the proposed all-in-one nanodevices were successfully applied to monitor intracellular microRNA-21 and telomerase expression levels. The flexible design can be extended to detect other cytoplasmic molecules and monitor related pathways by simply changing the sequences.