Electrochemical biosensor for DNA methyltransferase detection based on DpnI digestion triggering the formation of G-quadruplex DNAzymes

Liu, Pei; Liu, Min; Yin, Hunshun; Zhou, Yunlei; Ai, Shiyun

doi:10.1016/j.snb.2015.05.058

Cited by 26 publications

(3 citation statements)

References 27 publications

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“…142 However, traditional methods for DNA methyltransferase activity assay rely on radioisotope materials. 109 Therefore, radioisotope-free approaches have been developed, including electrochemical, [109][110][111][112][113][114][115][116][117][118][119] electrochemiluminescence, [120][121][122] photo-electrochemical, 123 fluorometric, [124][125][126][127][128][129][130][131][132][133][134][135] surface-enhanced Raman scattering (SERS), [136][137][138] and circular dichroism (CD) spectroscopy, 139 and surface plasmon resonance 140 based DNA methyltransferase measurement techniques have recently been proposed as summarized in Table 2 to realize simple, quick and easy monitoring.…”

Section: Dna Methyltransferase Activitymentioning

confidence: 99%

Microfluidic platforms for DNA methylation analysis

Kurita

Niwa

2016

Lab Chip

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In the field of genetics, epigenetics is the study of changes in gene expression without any change in DNA sequences. Chemical base modification in DNA by DNA methyltransferase, and specifically methylation, has been well studied as the main mechanism of epigenetics. Therefore, the determination of DNA methylation of, for example, 5'-methylcytosine in the CpG sequence in mammals has attracted attention because it should prove valuable in a wide range of research fields including diagnosis, drug discovery, and therapy. Methylated DNA bases and DNA methyltransferase activity are analyzed using conventional methods; however, these methods are time-consuming and require complex multiple operations. Therefore, new methods and devices for DNA methylation analysis are now being actively developed. Furthermore, microfluidic technology has also been applied to DNA methylation analysis because the microfluidic platform offers the promising advantage of making it possible to perform thousands of DNA methylation reactions in small reaction volumes, resulting in a high-throughput analysis with high sensitivity. This review discusses epigenetics and the microfluidic platforms developed for DNA methylation analysis.

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Section: Dna Methyltransferase Activitymentioning

confidence: 99%

Microfluidic platforms for DNA methylation analysis

Kurita

Niwa

2016

Lab Chip

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“…Based on methylation‐dependent endonuclease, Liu et al. reported an electrochemical biosensor to detect Dam MTase activity using a DNA probe richness of guanine and cofactor hemin combined with the methylation‐dependent endonuclease DpnⅠ .…”

Section: Introductionmentioning

confidence: 99%

A microchip electrophoretic assay for DNA methyltransferase activity based on methylation‐sensitive endonuclease DpnⅡ

Zhang

Zhu

et al. 2018

Electrophoresis

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DNA methylation is a significant epigenetic modification and the methods for the detection of DNA methyltransferase (MTase) activity are important due to aberrant methylation closely related to the occurrence of cancer. In this study, a simple and rapid microchip electrophoresis (ME) coupled with LED-induced fluorescence (LEDIF) method was presented for the detection of Dam MTase activity. This strategy was based on methylation-sensitive endonuclease DpnⅡ which could recognize the same specific site 5'-GATC-3' with Dam MTase in double-stranded DNA (dsDNA). The adenines in the specific site could be methylated by Dam MTase, then the special site could not be digested by DpnⅡ. Both methylated dsDNA and unmethylated dsDNA could be analyzed by ME-LEDIF after stained by SYBR gold. The results showed the fluorescence intensities of methylated dsDNA were directly proportional to Dam MTase activities in the range of 0.5-20 U/mL with a detection limit of 0.12 U/mL. Furthermore, the method could successfully be applied to evaluation experiments of Dam MTase inhibitors. The results confirmed the ME-LEDIF method is a promising approach for inhibitors screening of DNA MTase and development of anticancer drugs.

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“…Therefore, DNA methylation can be analyzed indirectly via assaying DNA MTase activity towards a target DNA. In recent years, much attention has been focused on developing electrochemical methods for measuring MTase activity(Li et al, 2015;Liu et al, 2015;Liu et al, 2016b). For example,Liu et al, (2011) reported an electrochemical assay for the detection of DNA methylation and the activity of MTase enzyme at specific CpG methylation sites.…”

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confidence: 99%

Electrochemical biosensing strategies for DNA methylation analysis

Hossain

Mahmudunnabi

Masud

et al. 2017

Biosensors and Bioelectronics

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DNA methylation is one of the key epigenetic modifications of DNA that results from the enzymatic addition of a methyl group at the fifth carbon of the cytosine base. It plays a crucial role in cellular development, genomic stability and gene expression. Aberrant DNA methylation is responsible for the pathogenesis of many diseases including cancers. Over the past several decades, many methodologies have been developed to detect DNA methylation. These methodologies range from classical molecular biology and optical approaches, such as bisulfite sequencing, microarrays, quantitative real-time PCR, colorimetry, Raman spectroscopy to the more recent electrochemical approaches. Among these, electrochemical approaches offer sensitive, simple, specific, rapid, and cost-effective analysis of DNA methylation. Additionally, electrochemical methods are highly amenable to miniaturization and possess the potential to be multiplexed. In recent years, several reviews have provided information on the detection strategies of DNA methylation. However, to date, there is no comprehensive evaluation of electrochemical DNA methylation detection strategies. Herein, we address the recent developments of electrochemical DNA methylation detection approaches. Furthermore, we highlight the major technical and biological challenges involved in these strategies and provide suggestions for the future direction of this important field.

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Electrochemical biosensor for DNA methyltransferase detection based on DpnI digestion triggering the formation of G-quadruplex DNAzymes

Cited by 26 publications

References 27 publications

Microfluidic platforms for DNA methylation analysis

Microfluidic platforms for DNA methylation analysis

A microchip electrophoretic assay for DNA methyltransferase activity based on methylation‐sensitive endonuclease DpnⅡ

Electrochemical biosensing strategies for DNA methylation analysis

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