DNA-based nanosensors for next-generation clinical diagnostics via detection of enzyme activity

Stougaard, Magnus; Ho, Yi‐Ping

doi:10.1586/14737159.2014.863151

Cited by 11 publications

(9 citation statements)

References 19 publications

(19 reference statements)

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“…AuNPs exhibit unique physicochemical properties, including their surface plasmon resonance (SPR), which relies on the interaction between an electromagnetic wave and free conduction electrons at the AuNPs’ surface, causing them to oscillate coherently in resonance with the frequency of visible light, resulting in strong electromagnetic fields. This phenomenon greatly enhances both the scattering and the absorption of light by the AuNP suitable for different biomedical applications 4 – 6 . AuNPs are also recognised by their PTT capacities, converting electromagnetic radiation into heat due to electron excitation and relaxation 5 , which has been used for thermal ablation of tumour cells.…”

Section: Introductionmentioning

confidence: 99%

Photothermal enhancement of chemotherapy in breast cancer by visible irradiation of Gold Nanoparticles

Mendes

Pedrosa

Lima

et al. 2017

Sci Rep

139

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Photothermal Therapy (PTT) impact in cancer therapy has been increasing due to the enhanced photothermal capabilities of a new generation of nanoscale photothermal agents. Among these nanoscale agents, gold nanoshells and nanorods have demonstrated optimal properties for translation of near infra-red radiation into heat at the site of interest. However, smaller spherical gold nanoparticles (AuNPs) are easier to produce, less toxic and show improved photoconversion capability that may profit from the irradiation in the visible via standard surgical green lasers. Here we show the efficient light-to-heat conversion of spherical 14 nm AuNPs irradiated in the visible region (at the surface plasmons resonance peak) and its application to selectively obliterate cancer cells. Using breast cancer as model, we show a synergistic interaction between heat (photoconversion at 530 nm) and cytotoxic action by doxorubicin with clear advantages to those of the individual therapy approaches.

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Section: Introductionmentioning

confidence: 99%

Photothermal enhancement of chemotherapy in breast cancer by visible irradiation of Gold Nanoparticles

Mendes

Pedrosa

Lima

et al. 2017

Sci Rep

139

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“…We have previously stressed the importance of using sensor systems resembling the natural substrates for the targeted enzymes as much as possible, i.e. , DNA-based sensor systems for DNA-modifying enzymes, since this will enable detection of differences in the DNA interaction of both enzymes and drugs [39]. In the REEAD assay such a DNA-based sensor system is exploited and is capable of detecting the cleavage-religation activity of hTopI at the single molecule level.…”

Section: Resultsmentioning

confidence: 99%

Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level

Proszek

Roy

Jakobsen

et al. 2014

Sensors

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Human topoisomerase I (hTopI) is an essential cellular enzyme. The enzyme is often upregulated in cancer cells, and it is a target for chemotherapeutic drugs of the camptothecin (CPT) family. Response to CPT-based treatment is dependent on hTopI activity, and reduction in activity, and mutations in hTopI have been reported to result in CPT resistance. Therefore, hTOPI gene copy number, mRNA level, protein amount, and enzyme activity have been studied to explain differences in cellular response to CPT. We show that Rolling Circle Enhanced Enzyme Activity Detection (REEAD), allowing measurement of hTopI cleavage-religation activity at the single molecule level, may be used to detect posttranslational enzymatic differences influencing CPT response. These differences cannot be detected by analysis of hTopI gene copy number, mRNA amount, or protein amount, and only become apparent upon measuring the activity of hTopI in the presence of CPT. Furthermore, we detected differences in the activity of the repair enzyme tyrosyl-DNA phosphodiesterase 1, which is involved in repair of hTopI-induced DNA damage. Since increased TDP1 activity can reduce cellular CPT sensitivity we suggest that a combined measurement of TDP1 activity and hTopI activity in presence of CPT will be the best determinant for CPT response.

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“…The RCPs obtained from the amplification in the qPCR machine were hybridized to the coupled primer and the microscopic pictures were acquired as previously reported [41]. Briefly, the slides were mounted with 2.5 µL Vectashield (Vector Laboratories, Burlingame, CA, USA) and a cover glass.…”

Section: Methodsmentioning

confidence: 99%

Optimized Detection of Plasmodium falciparum Topoisomerase I Enzyme Activity in a Complex Biological Sample by the Use of Molecular Beacons

Givskov

Kristoffersen

Vandsø

et al. 2016

Sensors

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The so-called Rolling Circle Amplification allows for amplification of circular DNA structures in a manner that can be detected in real-time using nucleotide-based molecular beacons that unfold upon recognition of the DNA product, which is being produced during the amplification process. The unfolding of the molecular beacons results in a fluorescence increase as the Rolling Circle Amplification proceeds. This can be measured in a fluorometer. In the current study, we have investigated the possibility of using two different molecular beacons to detect two distinct Rolling Circle Amplification reactions proceeding simultaneously and in the same reaction tube by measurement of fluorescence over time. We demonstrate the application of this fluorometric readout method, for automated and specific detection of the activity of the type IB topoisomerase from the malaria parasite Plasmodium falciparum in the presence of human cell extract containing the related topoisomerase I from humans. The obtained results point towards a future use of the presented assay setup for malaria diagnostics or drug screening purposes. In longer terms the method may be applied more broadly for real-time sensing of various Rolling Circle Amplification reactions.

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DNA-based nanosensors for next-generation clinical diagnostics via detection of enzyme activity

Cited by 11 publications

References 19 publications

Photothermal enhancement of chemotherapy in breast cancer by visible irradiation of Gold Nanoparticles

Photothermal enhancement of chemotherapy in breast cancer by visible irradiation of Gold Nanoparticles

Topoisomerase I as a Biomarker: Detection of Activity at the Single Molecule Level

Optimized Detection of Plasmodium falciparum Topoisomerase I Enzyme Activity in a Complex Biological Sample by the Use of Molecular Beacons

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