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DNAzyme–Metal–Organic Framework Two-Photon Nanoprobe for In situ Monitoring of Apoptosis-Associated Zn2+ in Living Cells and Tissues

原位 检出限 生物传感器 细胞生物学 纳米技术
作者
Xinxin Shi,Hong-Min Meng,Xin Geng,Lingbo Qu,Zhaohui Li
出处
期刊:ACS Sensors [American Chemical Society]
卷期号:5 (10): 3150-3157 被引量:42
标识
DOI:10.1021/acssensors.0c01271
摘要

Monitoring Zn2+ in living cells is critical for fully elucidating the biological process of apoptosis. However, the quantitative intracellular sensing of Zn2+ using DNAzyme remains challenging because of issues related to penetration of the signal through tissue, targeted cellular uptake and activation, and susceptibility toward enzymatic degradation. In this study, we developed a novel phosphate ion-activated DNAzyme–metal–organic frameworks (MOFs) nanoprobe for two-photon imaging of Zn2+ in living cells and tissues. The design of this nanoprobe involved the loading of a Zn2+-specific, RNA-cleaving DNAzyme onto the MOFs through strong coordination between the phosphonate O atoms of the DNAzyme backbone and Zr atoms in the MOFs. This coordination restrained the extracellular activity of DNAzyme; however, after cell entry, the DNAzyme was released from the MOFs through a competitive binding by the phosphate ions present at a high intracellular concentration. Following their release, the two-photon (TP) fluorophore-labeled substrate strands of DNAzyme were cleaved with the aid of Zn2+, which resulted in a strong fluorescence signal. The incorporation of a TP fluorophore into the nanoprobe facilitated near-infrared excitation, which allowed the highly sensitive and specific imaging of Zn2+ in living cells and tissues at greater depths than possible previously. The TP-DNAzyme-MOFs nanoprobe achieved a low detection limit of 3.53 nM, extraordinary selectivity toward Zn2+, and a tissue signal penetration of 120 μm. More importantly, this nanoprobe was successfully used to monitor cell apoptosis, and this application of the DNAzyme-MOFs probe holds great potential for future use in biological studies and medical diagnostics.
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