DNA methylation plays an important role in development process which contributes to genome stability and also regulates gene expression and gene silencing. Detection of genome regions with altered 5-methylcytosine distribution at a genome-wide scale is very important for early detection of gene silencing related diseases. In the present study as a continuation of studies on DNA methylation, the interactions between graphene quantum dots (GQDs) and unmethylated and methylated deoxyribonucleic acid (DNA) fragment were investigated. Based on above interaction a novel GQDs-DNA nanoassembly was developed. Two types of DNA including unmethylated and methylated sequences were interacted with GQDs and contributed to the formation of unmethylated and methylated nanoassemlies. Analysis of the interaction indicated that the GQDs could bind to DNA fragments and led to different fluorescence pattern in two different mechanisms and could provide an efficient biosensing platform for label free and sensitive fluorescent assay of DNA. The excitation and emission wavelengths of experiment were 380 and 480 nm respectively. Fluorescence intensity of unmethylated DNA concentration were detectable from methylated DNA in linear range from 10.0−10M to 10.0–6M and the detection limit was estimated at 7.3 × 10–11 M. Above interaction was not observed in methylated DNA, indicated of distinguished interaction effect. Herein we further showed that GQDs could induce B-DNA to A-DNA form in methylated structure of DNA. The methylation sensitivity of the experiment was also testified by methylation sensitive restriction process. It was assumed that the involvement of methylation alteration in DNA structure could alter not only mechanism of DNA/GDQs interaction but also helical structure of DNA.
DNA methylation has significant roles in gene regulation. DNA methyltransferase (MTase) enzyme characterizes DNA methylation and also induces an aberrant methylation pattern that is related to many diseases, especially cancers. Thus, it is required to develop a method to detect the DNA MTase activity. In this study, we developed a new sensitive and reliable method for methyltransferase activity assay by employing DNA-templated silver nanoclusters (DNA/Ag NCs) without using restriction enzymes. The Ag NCs have been utilized for the determination of M.SssI MTase activity and its inhibition. We designed an oligonucleotide probe which contained an inserted six-cytosine loop as Ag NCs formation template. The changes in fluorescence intensity were monitored to quantify the M.SssI activity. The fluorescence spectra showed a linear decrease in the range of 0.4 to 20 U/ml with a detection limit of 0.1 U/ml, which was significant compared with previous reports. The proposed method was applied successfully for demonstrating the Gentamicin effect as MTase inhibitor. The proposed method showed convenient reproducibility and sensitivity indicating its potential for the determination of methyltransferase activity.
Early detection of retroviruses including human T-cell lymphotropic virus and human immunodeficiency virus in the human body is indispensable to prevent retroviral infection propagation and improve clinical treatment. Until now, diverse techniques have been employed for the early detection of viruses. Traditional methods are timeconsuming, resource-intensive, and laborious performing. Therefore, designing and constructing a selective and sensitive diagnosis system to detect serious diseases is highly demanded. Genetic detection with high sensitivity has striking significance for the early detection and remedy of disparate pathogenic diseases. The nucleic acid biosensors are based on the identification of specific DNA sequences in biological samples. Nanotechnology has an important impact on the development of sensitive biosensors. Different kinds of nanomaterials include nanoparticles, nanoclusters, quantum dots, carbon nanotubes, nanocomposites, etc., with different properties have been used to improve the performance of biosensors. Recently, DNA nanobiosensors are developed to provide simple, fast, selective, low-cost, and sensitive detection of infectious diseases. In this paper, the research progresses of nano genosensors for the detection of HIV-1 and HTLV-1 viruses, based on electrochemical, optical, and photoelectrochemical platforms are overviewed.
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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