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[用于活细胞成像的荧光探针的开发]

[Development of a Fluorescence Probe for Live Cell Imaging].

作者信息

Shibata Aya

机构信息

Faculty of Engineering, Gifu University.

出版信息

Yakugaku Zasshi. 2017;137(11):1323-1337. doi: 10.1248/yakushi.17-00132.

DOI:10.1248/yakushi.17-00132
PMID:29093368
Abstract

Probes that detect specific biological materials are indispensable tools for deepening our understanding of various cellular phenomena. In live cell imaging, the probe must emit fluorescence only when a specific substance is detected. In this paper, we introduce a new probe we developed for live cell imaging. Glutathione S-transferase (GST) activity is higher in tumor cells than in normal cells and is involved in the development of resistance to various anticancer drugs. We previously reported the development of a general strategy for the synthesis of probes for detection of GST enzymes, including fluorogenic, bioluminogenic, and F-NMR probes. Arylsulfonyl groups were used as caging groups during probe design. The fluorogenic probes were successfully used to quantitate very low levels of GST activity in cell extracts and were also successfully applied to the imaging of microsomal MGST1 activity in living cells. The bioluminogenic and F-NMR probes were able to detect GST activity in Escherichia coli cells. Oligonucleotide-templated reactions are powerful tools for nucleic acid sensing. This strategy exploits the target strand as a template for two functionalized probes and provides a simple molecular mechanism for multiple turnover reactions. We developed a nucleophilic aromatic substitution reaction-triggered fluorescent probe. The probe completed its reaction within 30 s of initiation and amplified the fluorescence signal from 0.5 pM target oligonucleotide by 1500 fold under isothermal conditions. Additionally, we applied the oligonucleotide-templated reaction for molecular releasing and peptide detection.

摘要

检测特定生物材料的探针是深化我们对各种细胞现象理解的不可或缺的工具。在活细胞成像中,探针必须仅在检测到特定物质时才发出荧光。在本文中,我们介绍一种我们为活细胞成像开发的新探针。谷胱甘肽S-转移酶(GST)在肿瘤细胞中的活性高于正常细胞,并且参与对各种抗癌药物的耐药性发展。我们之前报道了一种用于合成检测GST酶的探针的通用策略,包括荧光探针、生物发光探针和F-核磁共振探针。在探针设计过程中,芳基磺酰基被用作封闭基团。荧光探针成功用于定量细胞提取物中极低水平的GST活性,并且还成功应用于活细胞中微粒体MGST1活性的成像。生物发光探针和F-核磁共振探针能够检测大肠杆菌细胞中的GST活性。寡核苷酸模板反应是核酸传感的强大工具。该策略利用靶链作为两个功能化探针的模板,并为多次周转反应提供了一种简单的分子机制。我们开发了一种亲核芳香取代反应触发的荧光探针。该探针在起始后30秒内完成反应,并在等温条件下将来自0.5 pM靶寡核苷酸的荧光信号放大了1500倍。此外,我们将寡核苷酸模板反应应用于分子释放和肽检测。

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1
[Development of a Fluorescence Probe for Live Cell Imaging].[用于活细胞成像的荧光探针的开发]
Yakugaku Zasshi. 2017;137(11):1323-1337. doi: 10.1248/yakushi.17-00132.
2
Design and synthesis of highly sensitive fluorogenic substrates for glutathione S-transferase and application for activity imaging in living cells.谷胱甘肽S-转移酶高灵敏度荧光底物的设计与合成及其在活细胞活性成像中的应用
J Am Chem Soc. 2008 Nov 5;130(44):14533-43. doi: 10.1021/ja802423n. Epub 2008 Oct 9.
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Reduction-triggered fluorescence probe for peptide-templated reactions.用于肽模板反应的还原触发荧光探针。
Chem Pharm Bull (Tokyo). 2009 Nov;57(11):1223-6. doi: 10.1248/cpb.57.1223.
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Very rapid DNA-templated reaction for efficient signal amplification and its steady-state kinetic analysis of the turnover cycle.非常快速的 DNA 模板反应,用于高效信号放大及其周转率的稳态动力学分析。
J Am Chem Soc. 2013 Sep 25;135(38):14172-8. doi: 10.1021/ja404743m. Epub 2013 Sep 9.
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Reduction-triggered fluorescent amplification probe for the detection of endogenous RNAs in living human cells.用于检测活人体细胞内源性 RNA 的还原触发荧光扩增探针。
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Optimized Fluorescent Probe for Specific Imaging of Glutathione S-Transferases in Living Cells and Mice.优化荧光探针用于活细胞和小鼠中谷胱甘肽 S-转移酶的特异性成像。
Chem Asian J. 2020 May 4;15(9):1464-1468. doi: 10.1002/asia.202000152. Epub 2020 Mar 30.
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A reduction-triggered fluorescence probe for sensing nucleic acids.一种用于检测核酸的还原触发荧光探针。
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Rapid miRNA imaging in cells using fluorogenic templated Staudinger reaction between PNA-based probes.利用基于肽核酸(PNA)的探针之间的荧光模板施陶丁格反应在细胞中进行快速微小RNA(miRNA)成像。
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Universal caging group for the in-cell detection of glutathione transferase applied to 19F NMR and bioluminogenic probes.用于细胞内谷胱甘肽转移酶检测的通用笼化基团,应用于19F核磁共振和生物发光探针。
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New, stronger nucleophiles for nucleic acid-templated chemistry: Synthesis and application in fluorescence detection of cellular RNA.用于核酸模板化学的新型更强亲核试剂:合成及其在细胞RNA荧光检测中的应用
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