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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. 在線等!!! |
版主 (文壇精英)
米米
金蟲 (小有名氣)

新蟲 (初入文壇)
| 恒量的Zn2+和Cu2+對活細胞的生理功能調節(jié)起重要作用。因此同時測定細胞中的Zn2+和Cu2+很有必要。本文,我們采用新型雙色熒光探針基于DNA酶同時對活細胞中的Zn2+和Cu2+成像。該探針由13nm金納米粒,特定的Zn2+和Cu2+DNA酶,及5′端用熒光基團標記、3′端用淬滅劑標記的底物鏈組成。熒光基團的熒光性同時被金納米粒和淬滅劑淬滅。熒光探針進去細胞之后,底物鏈會由于的Zn2+和Cu2+靶標的存在而斷裂,導致含熒光探針的DNA短片段的解離,從而產生了與Zn2+和Cu2+濃度及位置相關的熒光信號。納米探針展現了高特異性、核酸酶穩(wěn)定性及良好的生物相容性。此外,納米探針可實時實地地對細胞內的Zn2+和Cu2+進行監(jiān)測,從而提供靶標位置及濃度的相關信息,對進一步研究與Zn2+和Cu2+相關的細胞內物質及生理反應非常重要。本文提出的方法,可對活細胞中多金屬離子檢測,從而幫助我們更好地了解金屬離子在生物化學、分子生物學、細胞毒理學中的作用 |
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