DNAzyme self-assembled gold nanoparticles for determination of metal ions using fluorescence anisotropy assay

Yin, Bin‐Cheng; Zuo, Pengjian; Huo, Hao; Zhong, Xinhua; Ye, Bang‐Ce

doi:10.1016/j.ab.2010.02.014

Cited by 106 publications

(56 citation statements)

References 32 publications

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“…A number of fluorescent Pb 2+ sensors have been developed, which are based on the fluorescently labeled small organic chelators, 55,56 organic polymers, 57 peptides, 58 and functional nucleic acid. 3,9 The detection limit of our nonnanoparticle-dependent FA approach (100 pM) is lower than that of nanoparticle-dependent FA approaches (LOD, 1 nM,) 18 and other fluorescence intensity-based homogeneous analysis (LOD, 3.7 nM). 38 Our approach for detection of free Pb 2+ has several advantages.…”

Section: ■ Results and Discussionmentioning

confidence: 79%

Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity

Zhang

Zhao

et al. 2015

Anal. Chem.

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Fluorescence anisotropy is a homogeneous, sensitive, ratiometric, and real-time analytical technology. However, it is a great challenge to produce a large fluorescence anisotropy change upon the presence of target small molecules without nanoparticles-dependent amplification. This work reports a nanoparticle-free and multiple G-enhanced fluorescence anisotropy assay for detection of DNAzyme activity. A Pb 2+ -dependent GR-5 DNAzyme was used as a model. We hybridized the rA-cleavable substrate strand containing a TMR label at the 5′-end with the DNAzyme strand containing an extended three G bases at the 3′-end. By this design, we demonstrate that both fluorescence quenching and the enhanced DNAzyme activity contribute to a Pb 2+ -induced large fluorescence anisotropy change (|Δr| = 0.168). The limit of detection for Pb 2+ is estimated to be about 100 pM with a dynamic range from 200 pM to 100 nM. The interference from the other nine divalent metal ions of 1000-times excess amount is negligible. Moreover, we show an extended assay for evaluation of the interactions of Pb 2+ with cysteine and glutathione by the detection of GR5 DNAzyme activity. Collectively, we developed a novel fluorescence anisotropy amplification assay, enabling us to detect DNAzyme activity and associated cofactors and inhibitors and to characterize the Pb 2+ -chelation capability of free thiols.O ne class of functional nucleic acids with catalytic activity is DNAzymes, which have potential applications in diagnosis, therapeutics, and bioanalysis. 1−4 Various DNAzymes have been in vitro selected to catalyze a wide range of reactions, such as RNA cleavage, DNA cleavage, RNA ligation, DNA ligation, DNA phosphorylation, cleavage of the phosphoramidate bond, and porphyrin metalation. 5,6 One type of DNAzymes can cleave RNA. These RNA-cleaving DNAzymes are composed of a DNA enzyme strand and a DNA substrate strand containing a single RNA linkage (rA), which serves as the cleavage site. 7 This class of DNAzymes has been applied in nucleic acid detection, 8 metal ions sensing, 3,4,6,7,9−11 and molecular logic gate constructing. 4,12,13 The enzymatic activities play central roles in any extended analytical applications of DNAzymes. A number of biochemical and biophysical methods have been carried out for measuring DNAzyme activities, especially for RNA-cleaving DNAzymes. These methods include polyacrylamide gel electrophoresis (PAGE), 14 MALDI-TOF mass spectrometry, 15 fluorescence resonance energy transfer (FRET), 16,17 and fluorescence anisotropy (FA). 18−22 Among known techniques for detection of DNAzyme activity, FA has a number of advantages. It is a simple, realtime, and truly homogeneous analysis. Moreover, it does not require the separation of substrates and the cleaved products. This technique also presents several other attractive features, such as the requirement of only one fluorescent label and relative insensitivity to the photobleaching of the fluorophore and to instrument parameters due to its ratiometric nature. In fact, F...

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Section: ■ Results and Discussionmentioning

confidence: 79%

Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity

Zhang

Zhao

et al. 2015

Anal. Chem.

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“…In addition to fluorescence quenching, AuNPs could serve as fluorescence anisotropy generators for the coated fluorophores due to the large size compared to free fluorophores and DNA. Based on this principle, a fluorescence anisotropy sensor was constructed using DNAzyme-functionalized AuNPs for the detection of Cu 2+ and Pb 2+ [259]. Finally, DNAzyme-functionalized AuNPs were also incorporated with electrochemically active substances for biosensor applications.…”

Section: Nucleic Acid Cleaving/ligating Dnazymes and Aunps For Biosenmentioning

confidence: 99%

DNAzyme-Functionalized Gold Nanoparticles for Biosensing

Xiang

Tan

et al. 2013

Advances in Biochemical Engineering/Biotechnology

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Recent progress in using DNAzyme-functionalized gold nanoparticles (AuNPs) for biosensing is summarized in this chapter. A variety of methods, including those for attaching DNA on AuNPs, detecting metal ions and small molecules by DNAzyme-functionalized AuNPs, and intracellular applications of DNAzyme-functionalized AuNPs are discussed. DNAzyme-functionalized AuNPs will increasingly play more important roles in biosensing and many other multidisciplinary applications. This chapter covers the recent advancement in biosensing applications of DNAzyme-functionalized gold nanoparticles, including the detection of metal ions, small molecules, and intracellular imaging.

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“…Of these, [8][9][10][11][12][13][14][15][16][17] DNAzyme has attracted much more attention. They have been extensively explored for Pb 2+ detection via fluorescence [12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29], fluorescence anisotropy [30], colorimetry [31][32][33][34], electrochemistry [35][36][37][38], electrochemiluminescence [39], dynamic light scattering [40,41], and surface enhanced Raman scattering [42].…”

Section: Introductionmentioning

confidence: 99%

A sensitive fluorescence anisotropy method for detection of lead (II) ion by a G-quadruplex-inducible DNA aptamer

Zhang

Meng

et al. 2014

Analytica Chimica Acta

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h i g h l i g h t s• A fluorescence anisotropy approach for detection of Pb 2+ was developed.• The strategy was based on bindinginduced allosteric conformational change of aptamer probe.• The sensing mechanism was established by testing the photoinduced electron transfer interaction. in homogeneous solution by a G-rich thrombin binding aptamer (TBA). The TBA labeled with 6-carboxytetramethylrhodamine (TMR) at the seventh thymine nucleotide was used as a fluorescent probe for signaling Pb 2+ . It was found that the aptamer probe had a high FA in the absence of Pb 2+ . This is because the rotation of TMR is restricted by intramolecular interaction with the adjacent guanine bases, which results in photoinduced electron transfer (PET). When the aptamer probe binds to Pb 2+ to form G-quadruplex, the intramolecular interaction should be eliminated, resulting in faster rotation of the fluorophore TMR in solution. Therefore, FA of aptamer probe is expected to decrease significantly upon binding to Pb 2+ . Indeed, we observed a decrease in FA of aptamer probe upon Pb 2+ binding. Circular dichroism, fluorescence spectra, and fluorescence lifetime measurement were used to verify the reliability and reasonability of the sensing mechanism. By monitoring the FA change of the aptamer probe, we were able to real-time detect binding between the TBA probe and Pb 2+ . Moreover, the aptamer probe was exploited as a recognition element for quantification of Pb 2+ in homogeneous solution. The change in FA showed a linear response to Pb 2+ from 10 nM to 2.0 M, with 1.0 nM limit of detection. In addition, this sensing system exhibited good selectivity for Pb 2+ over other metal ions. The method is simple, quick and inherits the advantages of aptamer and FA.

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DNAzyme self-assembled gold nanoparticles for determination of metal ions using fluorescence anisotropy assay

Cited by 106 publications

References 32 publications

Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity

Nanoparticles-Free Fluorescence Anisotropy Amplification Assay for Detection of RNA Nucleotide-Cleaving DNAzyme Activity

DNAzyme-Functionalized Gold Nanoparticles for Biosensing

A sensitive fluorescence anisotropy method for detection of lead (II) ion by a G-quadruplex-inducible DNA aptamer

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