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Trace Zn2+ and Cu2+ in living cells play important roles in the regulation of biological function. It is significant to simultaneously detect the cellular Zn2+ and Cu2+. Here, we present a novel two-color fluorescence nanoprobe based on the DNAzymes for simultaneous imaging of Zn2+ and Cu2+ in living cells. The probe consists of a 13 nm gold nanoparticle, DNAzymes that are specific for Zn2+ and Cu2+, and the substrate strands labeled with fluorophores at the 5′ end and quenchers at the 3′ end. The fluorescence of the fluoreophores is quenched both by the gold nanoparticle and the quencher. After the nanoprobes are transferred into the cells, the substrate strands would be cleaved in the presence of the Zn2+ and Cu2+ target, resulting in disassociation of the shorter DNA fragments containing fluorophores, which produce fluorescence signals correlated with the location and concentration of the Zn2+ and Cu2+. The nanoprobe exhibits high specificity, nuclease stability, and good biocompatibility. Moreover, the nanoprobe can simultaneously monitor the cellular Zn2+ and Cu2+ with an on-site manner, providing the information on localization and concentration of targets, which is significant to further research the Zn2+- and Cu2+-relative cellular events and biological process. The proposed method has shown great potential in the detection of multiple metal ions in living cells, which may help us to better understand the function of metal ions in the fields of biochemistry, molecular biology, and cellular toxicology. 在线等!!! |
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2楼2015-06-14 16:49:52
fjtony163
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| 恒量的Zn2+和Cu2+对活细胞的生理功能调节起重要作用。因此同时测定细胞中的Zn2+和Cu2+很有必要。本文，我们采用新型双色荧光探针基于DNA酶同时对活细胞中的Zn2+和Cu2+成像。该探针由13nm金纳米粒，特定的Zn2+和Cu2+DNA酶，及5′端用荧光基团标记、3′端用淬灭剂标记的底物链组成。荧光基团的荧光性同时被金纳米粒和淬灭剂淬灭。荧光探针进去细胞之后，底物链会由于的Zn2+和Cu2+靶标的存在而断裂，导致含荧光探针的DNA短片段的解离，从而产生了与Zn2+和Cu2+浓度及位置相关的荧光信号。纳米探针展现了高特异性、核酸酶稳定性及良好的生物相容性。此外，纳米探针可实时实地地对细胞内的Zn2+和Cu2+进行监测，从而提供靶标位置及浓度的相关信息，对进一步研究与Zn2+和Cu2+相关的细胞内物质及生理反应非常重要。本文提出的方法，可对活细胞中多金属离子检测，从而帮助我们更好地了解金属离子在生物化学、分子生物学、细胞毒理学中的作用 |
5楼2015-07-07 12:29:56
