Imaging Endogenous Metal Ions in Living Cells Using a DNAzyme–Catalytic Hairpin Assembly Probe,Angewandte Chemie International Edition

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Imaging Endogenous Metal Ions in Living Cells Using a DNAzyme–Catalytic Hairpin Assembly Probe
Angewandte Chemie International Edition ( IF 16.1 ) Pub Date : 2017-06-23 03:41:34 , DOI: 10.1002/anie.201703540
Zhenkun Wu _{1,

2} , Huanhuan Fan _{1,

2} , Nitya Sai Reddy Satyavolu ₂ , WenJing Wang _{2,

3} , Ryan Lake ₂ , Jian-Hui Jiang ₁ , Yi Lu ₂

Affiliation

DNAzymes are a promising platform for metal ion detection, and a few DNAzyme-based sensors have been reported to detect metal ions inside cells. However, these methods required an influx of metal ions to increase their concentrations for detection. To address this major issue, the design of a catalytic hairpin assembly (CHA) reaction to amplify the signal from photocaged Na⁺-specific DNAzyme to detect endogenous Na⁺ inside cells is reported. Upon light activation and in the presence of Na⁺, the NaA43 DNAzyme cleaves its substrate strand and releases a product strand, which becomes an initiator that trigger the subsequent CHA amplification reaction. This strategy allows detection of endogenous Na⁺ inside cells, which has been demonstrated by both fluorescent imaging of individual cells and flow cytometry of the whole cell population. This method can be generally applied to detect other endogenous metal ions and thus contribute to deeper understanding of the role of metal ions in biological systems.

中文翻译：

使用DNAzyme催化发夹组装探针成像活细胞中的内源性金属离子

DNAzyme是用于金属离子检测的有前途的平台，据报道，一些基于DNAzyme的传感器可检测细胞内的金属离子。但是，这些方法需要大量金属离子以增加其浓度以进行检测。为了解决这个主要问题，已经报道了设计催化发夹装配（CHA）反应以放大来自光笼罩的Na ⁺特异性DNA核酶的信号以检测细胞内的内源性Na ^+的设计。光激活后，在Na ⁺存在下，NaA43 DNAzyme裂解其底物链并释放产物链，该产物链成为引发随后CHA扩增反应的引发剂。这种策略可以检测内源性Na ⁺单个细胞的荧光成像和整个细胞群体的流式细胞术都已证明了这一点。该方法通常可用于检测其他内源性金属离子，因此有助于更深入地了解金属离子在生物系统中的作用。

更新日期：2017-06-24

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