Triplex-Functionalized DNA Tetrahedral Nanoprobe for Imaging of Intracellular pH and Tumor-Related Messenger RNA
Triplex-Functionalized DNA Tetrahedral Nanoprobe for Imaging of Intracellular pH and Tumor-Related Messenger RNA
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用于细胞内 pH 和肿瘤相关信使 RNA 成像的三重功能化 DNA 四面体纳米探针
DOI：
10.1021/acs.analchem.9b03659
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发表时间：
2019
影响因子：
7.4
通讯作者：
Ronghua Yang
中科院分区：
文献类型：
--
作者：
Cong Zhu;Jinfeng Yang;Jing Zheng;Shiya Chen;Fujian Huang;Ronghua Yang
本文提出了一种新的三链体功能化的DNA四面体纳米探针，用于监测活细胞中的pH和信使RNA（mRNA）。与传统的基于DNA四面体的纳米探针不同，DNA三链体被用作重要的构象转换元件。受肿瘤细胞外低pH环境的启发，以mRNA为靶点的H1和H2通过Hoogsteen键合稳定地组装在DNA四面体的延伸短发夹探针上，形成DNA三链体。由于高细胞内pH和靶mRNA的存在，在DNA三链体解离释放的H1和H2之间触发杂交链反应（HCR），并且所产生的长双链DNA激活Fo李斯特共振能量转移（FRET）信号，即使在非常低的含量下也指示靶mRNA表达。通过结合DNA三链体结构（pH响应）和HCR（信号放大）的特点，可以实现细胞内pH和肿瘤相关mRNA的灵敏成像。作为进一步的应用，在人乳腺癌细胞中成功地实现了细胞内pH和mRNA在“依赖性”途径凋亡过程中的动态成像，这表明我们提出的纳米探针在疾病的早期诊断和治疗中具有巨大的潜力。
A new triplex-functionalized DNA tetrahedral nanoprobe is proposed herein for monitoring pH and messenger RNA (mRNA) in living cells. Different from traditional DNA tetrahedron-based nanoprobes, DNA triplex was employed to serve as important conformational conversion elements. Inspired by the low extracellular pH in tumor cells, the mRNA-targeted H1 and H2 were stably assembled on the extended short hairpin probes of DNA tetrahedron via Hoogsteen bonding to form DNA triplex. Due to the high intracellular pH and presence of target mRNA, hybridization chain reaction (HCR) was triggered between H1 and H2 which were released from the dissociation of DNA triplex, and the generated long double-stranded DNA activated a Föster resonance energy transfer (FRET) signal indicating target mRNA expression even at very low contents. By combining the distinguishing feature of DNA triplex structure (pH-responsive) and HCR (signal amplification), sensitive imaging of intracellular pH and tumor-related mRNA can be realized. As a further application, dynamic imaging of intracellular pH and mRNA during “mitochondria-dependent” pathway apoptosis was successfully achieved in human breast cancer cells, which indicated huge potential of our proposed nanoprobe in early diagnosis and treatment of diseases.