Sensitive and selective real-time electrochemical monitoring of DNA repair

McWilliams, Marc A.; Anka, Fadwa H.; Balkus, Kenneth J.; Slinker, Jason D.

doi:10.1016/j.bios.2013.11.034

Cited by 52 publications

(27 citation statements)

References 47 publications

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“…Notably, we showed that these sensors selectively followed repair of 8-oxoguanine-modifed DNA by FPG in our previous work (McWilliams et al 2014). This is important as glycosylases, such as FPG, bind to DNA nonspecifically but excise bases with high damage specificity, requiring sensors that transduce base excision, rather than DNA binding, to distinguish activity over controls.…”

Section: Resultsmentioning

confidence: 83%

“…Given the anticipated generation of H 2 O 2 of ~0.3 mM, this change is consistent with responses from stand-alone Fenton reaction of higher H 2 O 2 concentrations. The change observed in drug-free electrodes is associated with nonspecific binding by FPG (McWilliams et al 2014). Additional controls were performed with β-lap present, but with other factors of the futile redox cycle removed.…”

Section: Resultsmentioning

confidence: 99%

“…Changes in the DNA structure, such as removal of a damaged base by a repair enzyme, lowered the voltammetry peak. We recently reported such an assay that detects base excision repair (BER) sensitively (down to nanograms of the enzyme), selectively (against undamaged controls), and in real-time with the natural substrate (McWilliams et al 2014). Here, we demonstrate the observation of drug-specific damage associated with therapeutic levels of the anticancer agent, ß-lapachone (ß-lap).…”

Section: Introductionmentioning

confidence: 99%

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Using DNA devices to track anticancer drug activity

Kahanda

Chakrabarti

McWilliams

et al. 2016

Biosensors and Bioelectronics

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It is beneficial to develop systems that reproduce complex reactions of biological systems while maintaining control over specific factors involved in such processes. We demonstrated a DNA device for following the repair of DNA damage produced by a redox-cycling anticancer drug, beta-lapachone (β-lap). These chips supported ß-lap-induced biological redox cycle and tracked subsequent DNA damage repair activity with redox-modified DNA monolayers on gold. We observed drug-specific changes in square wave voltammetry from these chips at therapeutic ß-lap concentrations of high statistical significance over drug-free control. We also demonstrated a high correlation of this change with the specific ß-lap-induced redox cycle using rational controls. The concentration dependence of ß-lap revealed significant signal changes at levels of high clinical significance as well as sensitivity to sub-lethal levels of ß-lap. Catalase, an enzyme decomposing peroxide, was found to suppress DNA damage at a NQO1/catalase ratio found in healthy cells, but was clearly overcome at a higher NQO1/catalase ratio consistent with cancer cells. We found that it was necessary to reproduce key features of the cellular environment to observe this activity. Thus, this chip-based platform enabled tracking of ß-lap-induced DNA damage repair when biological criteria were met, providing a unique synthetic platform for uncovering activity normally confined to inside cells.

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Section: Resultsmentioning

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Using DNA devices to track anticancer drug activity

Kahanda

Chakrabarti

McWilliams

et al. 2016

Biosensors and Bioelectronics

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“…In our previous work, the photoelectrochemical sensor was used briefly for the detection of 8-oxodGuo excision by FPG (Zhang et al, 2012). More recently, real-time electrochemical monitoring of 8-oxodGuo and uracil repair by DNA glycosylases were achieved (McWilliams et al, 2014). With these in mind, in the present work we developed a DNA biosensor for the detection of 8-oxodGuo lesion repair by FPG enzyme.…”

Section: Introductionmentioning

confidence: 99%

An electrochemiluminescence biosensor for 8-oxo-7,8-dihydro-2′-deoxyguanosine quantification and DNA repair enzyme activity analysis using a novel bifunctional probe

Yang

Zhang

et al. 2015

Biosensors and Bioelectronics

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a b s t r a c tA new electrochemiluminescence (ECL) sensor was developed for 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodGuo) quantification and Escherichia coli formamidopyrimidine-DNA glycosylase (FPG) activity assay. The sensor employed a novel spermine conjugated ruthenium tris-(bipyridine) derivative (spermine-Ru) which binds specifically with 8-oxodGuo through a one-step reaction and also acts as an ECL signal reporter. In the sensor, an 8-oxodGuo-containing ds-DNA film was first immobilized on a gold electrode by self-assembly. The DNA film was then incubated with spermine-Ru under oxidative condition for 8-oxodGuo labeling. The ECL intensity was found to correlate with the amount of 8-oxodGuo on the surface and the detection limit was estimated to be about 1 lesion in 500 DNA bases. Addition of FPG resulted in some loss of the signal due to the excision of 8-oxodGuo by the enzyme. An inverse relationship between ECL intensity and FPG concentration was observed in a range from 0 to 4.0 U/mL, demonstrating that this sensor could be used for FPG activity assay. A number of metal ions were screened by the sensor for their inhibition effect on FPG activity. Among them, Hg 2 þ and methyl Hg(II)shown very potent inhibition, with IC 50 values of 4.04 mM and 4.34 nM respectively. The result may suggest that interference on the DNA repair system could be another mechanism for the high toxicity of MeHg.

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“…We have demonstrated devices that follow DNA damage repair in real time, with a convenient, low-cost package (see Figure 1). 1 In this device, DNA is bound to the circular electrodes of multielectrode chips, and a redox probe at the top of the DNA reports charge transfer through it. DNA is the natural recognition element not only for the binding of repair proteins but also for their repair activity, and it can be synthesized with or without damage/lesion sites to establish controls.…”

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confidence: 99%