Gold nanorods<b>-</b>based FRET assay for ultrasensitive detection of DNA methylation and DNA methyltransferase activity

Wang, Gang Lin; Luo, Hong Qun; Li, Nian Bing

doi:10.1039/c4an00206g

Cited by 21 publications

(17 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the regression equation, x value represents the concentration of Dam MTase (U/mL) and the y value is fluorescence intensity of methylated dsDNA (mV). The detection limit of the ME method is higher or similar to previous reported methods, which suggests that this method is efficient to detect the activity of Dam MTase .The RSDs ( n = 3) of the migration time and fluorescence intensities were 1.7 and 2.5%, respectively. These data indicate that the ME method for Dam MTase activity assay has good reproducibility and high precision.…”

Section: Resultssupporting

confidence: 71%

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

Zhang

Zhu

et al. 2018

Electrophoresis

View full text Add to dashboard Cite

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.

show abstract

Section: Resultssupporting

confidence: 71%

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

Zhang

Zhu

et al. 2018

Electrophoresis

View full text Add to dashboard Cite

show abstract

“…Also, many reports used oligonucleotide cleavage based methods for MTase enzyme detection. 41,42 It is worth noting that we reported the cleavage free MTase enzyme detection for the rst time. The induction of a methyl group on the DNA structure may contribute to the recovery of the quenched FAM uorophore through its electron donating activity.…”

Section: 3 Detection Of Dna Methylation Induced By Msssi Mtasementioning

confidence: 94%

Sensitive detection of methylated DNA and methyltransferase activity based on the lighting up of FAM-labeled DNA quenched fluorescence by gold nanoparticles

et al. 2019

View full text Add to dashboard Cite

show abstract

“…In the increasing popularity of nanomaterials, some groups incorporated nanoparticles such as silver nanoclusters 61 , carbon nanotubes 73 and gold nanorods 74 into the assays based on the strong affinity each type of nanomaterials has with either a particular base present in the substrate DNA, with ss-DNA or ds-DNA. Consequently, the review below seeks to give an overview yet detailed explanation of these novel assays.…”

Section: Dna Mtase Activity Assaysmentioning

confidence: 99%

“…In a recent report, FRET was employed between gold nanorods (AuNRs) and fluorescein FAM-tagged substrate ds-DNA by Wang and colleagues 74 . Due to high affinity or aggregation formation between the AuNRs and the substrate ds-DNA, the substrate ds-DNA thus forms a hybrid with the FAM-tagged DNA which will have electrostatic interaction with the AuNRs.…”

Section: Dna Mtase Activity Assaysmentioning

confidence: 99%

DNA Methyltransferase Activity Assays: Advances and Challenges

Poh¹,

Wee²,

Gao³

2016

Theranostics

View full text Add to dashboard Cite

DNA methyltransferases (MTases), a family of enzymes that catalyse the methylation of DNA, have a profound effect on gene regulation. A large body of evidence has indicated that DNA MTase is potentially a predictive biomarker closely associated with genetic disorders and genetic diseases like cancer. Given the attention bestowed onto DNA MTases in molecular biology and medicine, highly sensitive detection of DNA MTase activity is essential in determining gene regulation, epigenetic modification, clinical diagnosis and therapeutics. Conventional techniques such as isotope labelling are effective, but they often require laborious sample preparation, isotope labelling, sophisticated equipment and large amounts of DNA, rendering them unsuitable for uses at point-of-care. Simple, portable, highly sensitive and low-cost assays are urgently needed for DNA MTase activity screening. In most recent technological advances, many alternative DNA MTase activity assays such as fluorescent, electrochemical, colorimetric and chemiluminescent assays have been proposed. In addition, many of them are coupled with nanomaterials and/or enzymes to significantly enhance their sensitivity. Herein we review the progress in the development of DNA MTase activity assays with an emphasis on assay mechanism and performance with some discussion on challenges and perspectives. It is hoped that this article will provide a broad coverage of DNA MTase activity assays and their latest developments and open new perspectives toward the development of DNA MTase activity assays with much improved performance for uses in molecular biology and clinical practice.

show abstract

Gold nanorods-based FRET assay for ultrasensitive detection of DNA methylation and DNA methyltransferase activity

Cited by 21 publications

References 36 publications

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

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

Sensitive detection of methylated DNA and methyltransferase activity based on the lighting up of FAM-labeled DNA quenched fluorescence by gold nanoparticles

DNA Methyltransferase Activity Assays: Advances and Challenges

Contact Info

Product

Resources

About