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A Two-photon DNAzyme-Gold Nanoparticle Probe for Imaging Intracellular Metal Ions Chan Yang, Xia Yin, Shuang-Yan Huan, Lanlan Chen, XiaoXiao Hu, Mengyi Xiong, Kun Chen, and Xiao-Bing Zhang Anal. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.analchem.7b04171 • Publication Date (Web): 07 Feb 2018 Downloaded from http://pubs.acs.org on February 8, 2018
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Analytical Chemistry
A Two-photon DNAzyme-Gold Nanoparticle Probe for Imaging Intracellular Metal Ions Chan Yang†, Xia Yin†, Shuang-Yan Huan*†, Lanlan Chen*‡, Xiao-Xiao Hu†, Meng-yi Xiong†, Kun Chen†, Xiao-Bing Zhang*† †
Molecular Science and Biomedicine Laboratory, College of Chemistry and Chemical
Engineering and College of Biology, State Key Laboratory of Chemo/Biosensing and Chemometrics, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha 410082. ‡
Shandong Provincial Key Laboratory of Detection Technology for Tumour Markers,
College of Chemistry and Chemical Engineering, Linyi University, Linyi, Shandong 276005, P. R. China *To whom correspondence should be addressed. E-mail:
[email protected] [email protected] [email protected] 1
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ABSTRACT: RNA-cleaving DNAzymes have been demonstrated as a promising platform for sensing metal ions. However, the poor biological imaging performance of RNA cleaving DNAzyme-based fluorescent probes has limited their intracellular applications. Compared with traditional one photon fluorescence imaging, two photon (TP) fluorescent probes have shown advantages such as increased penetration depth, lower tissue autofluorescence, and reduced photo damage. Herein, for the first time, we
developed
a
RNA-cleaving
DNAzyme-based
TP
imaging
probe
(TP-8-17ES-AuNP) for Zn2+ detection in living cells by modifying Zn2+-specific DNAzyme (8-17) with TP fluorophore (TP-8-17ES), and using gold nanoparticles (AuNPs) for intracellular delivery. The modified TP-8-17ES exhibits good two photon properties and excellent photostability. For the TP-8-17ES-AuNP, in the absence of Zn2+, the TP fluorophore is quenched by both AuNPs and the molecular quencher. Only in the presence of Zn2+, the DNAzyme cleaves the TP fluorophore labeled substrate strand, resulting in fluorescence enhancement and TP imaging. Such probe shows remarkable selectivity of Zn2+ over other metal ions existed in biological environment. Benefitting from the labeled TP fluorophore, the near-infrared (NIR) excited probe has the capability of TP imaging of Zn2+ in living cells and tissue with a deep tissue penetration up to 160 µm. This method can be generally applied to detect other metal ions in biological systems under TP imaging with higher tissue penetration ability and lower phototoxicity.
INTRODUCTION: 2
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Analytical Chemistry
Metal ions play vital roles in regulating physiological and pathological processes1. Thus, gaining detail information about the location and the concentration of metal ions in living cells is of significant importance2-7. Numerous efforts have been made to develop fluorescent probes for metal ions detection. Among them, RNA-cleaving DNAzyme-based sensors have attracted more and more attention, as RNA-cleaving DNAzymes have high specificity for a specific metal ion resulted from the complicated in vitro selection process8-14. Moreover, RNA-cleaving DNAzymes have advantages of being biocompatible, water-soluble, low toxicity and easy to synthesis and functionalization, making RNA-cleaving DNAzymes to be a more general platform to develop sensors for as many metal ions as possible15-19. Nevertheless, with the difficulties of RNA-cleaving DNAzymes delivery into desired locations and maintaining its catalytic activity in living cells, only few works have been reported on RNA-cleaving DNAzyme-based sensors for intracellular metal ions detection in the early stage20-26. In 2013, Lu and coworkers reported the first gold nanoparticles (AuNPs)-based DNAzyme sensor for detecting UO22+ in living cells20. Li et al.21 developed a two-color AuNPs-DNAzyme fluorescence probe simultaneously tracing intracellular Zn2+ and Cu2+. However, up to now, most reported RNA-cleaving DNAzyme-based fluorescence probes are modified by one-photon (OP) fluorophore excited by short wavelength, resulting in poor biological imaging with shallow tissue penetration depth (
