Sensitive detection of DNA methyltransferase activity by transcription-mediated duplex-specific nuclease-assisted cyclic signal amplification

Zhang, Yan; Xu, Wenjing; Zeng, Ya-ping; Zhang, Chunyang

doi:10.1039/c5cc05922d

Cited by 29 publications

(16 citation statements)

References 42 publications

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“…In this study IC50 has been found to be 3.1 µM which indicates that this is a promising method to screen the inhibitor effects of drugs on MTase [56,57].…”

Section: Fig 3 Msssi Methyltransferase Inhibition Assaymentioning

confidence: 66%

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A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters

et al. 2018

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DNA methylation catalyzed by DNA methyl transferase (MTase) is a significant epigenetic process for modulating gene expression. Abnormal levels of DNA MTase enzyme have been regarded as a cancer biomarker or a sign of bacterial diseases. We developed a novel colorimetric method to assay M.SssI MTase activity employing peroxidase-like activity of DNA template Ag/Pt NCs without using restriction enzymes. Based on inhibiting the peroxidase reaction that occurred in the TMB-HO system, in the presence of MTase, a highly sensitive and selective colorimetric biosensor was fabricated with a detection limit (LOD) of 0.05 U/mL and a linear range from 0.5 to 10 U/mL. The changes in absorption intensity were monitored to quantify the M.SssI activity. This strategy had a high selectivity over other proteins. Furthermore, it is also demonstrated that this method can be used for the evaluation and screening of inhibitors for DNA MTase.

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“…In this study IC50 has been found to be 3.1 µM which indicates that this is a promising method to screen the inhibitor effects of drugs on MTase [56,57].…”

Section: Fig 3 Msssi Methyltransferase Inhibition Assaymentioning

confidence: 66%

“…To check the validity of our method in screening the inhibitors of M.SssI, we have employed Gentamicin as an inhibitor model [56]. We have evaluated the inhibitor effects by incubating DNA-Ag/Pt NCs and M.SssI MTase and SAM with different doses of Gentamicin (Fig.…”

Section: Fig 3 Msssi Methyltransferase Inhibition Assaymentioning

confidence: 99%

A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters

et al. 2018

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“…DSN is widely used in fulllength enriched cDNA normalization, 18,19 construction of normalized RNA-seq libraries for next generation sequencing 20,21 and molecular biology. [22][23][24] Very recently, based on excellent selectivity in digestion of DSN, the Ye group has established a new signal-amplifying mechanism, which was duplex-specific nuclease signal amplification (DSNSA), for highly sensitive detection of miRNA. 25 From then on, several miRNA detection methods based on DSN have been developed.…”

Section: Introductionmentioning

confidence: 99%

A simple molecular beacon with duplex-specific nuclease amplification for detection of microRNA

Zhang

Zhao

et al. 2016

Analyst

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MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene activity, promoting or inhibiting cell proliferation, migration and apoptosis. Abnormal expression of miRNAs is associated with many diseases. Therefore, it is essential to establish a simple, rapid and sensitive miRNA detection method. In this paper, based on a simple molecular beacon (MB) and duplex-specific nuclease (DSN), we developed a target recycling amplification method for miRNA detection. By controlling the number of stem bases to 5, the MB probe used in this method can be prevented from hydrolysis by DSN without special modification. This assay is direct and simple to quantitatively detect miRNA with high sensitivity and specificity. The MB probe design provides a new strategy for nuclease-based amplification reaction.

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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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Sensitive detection of DNA methyltransferase activity by transcription-mediated duplex-specific nuclease-assisted cyclic signal amplification

Cited by 29 publications

References 42 publications

A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters

A colorimetric assay of DNA methyltransferase activity based on peroxidase mimicking of DNA template Ag/Pt bimetallic nanoclusters

A simple molecular beacon with duplex-specific nuclease amplification for detection of microRNA

Microfluidic platforms for DNA methylation analysis

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