Nano-graphene oxide as a novel platform for monitoring the effect of LNA modification in nucleic acid interactions (original) (raw)
Related papers
Graphene oxide has gained significant attention due to its exceptional physical properties at biological interfaces. It has extraordinary quenching, fast adsorption and desorption properties that are suitable for detection of molecular interactions in nucleic acids. Here we studied the interaction between locked nucleic acid (LNA) modified oligonucleotides and its complementary miR-10b DNA analog. We demonstrate that LNA modification does not alter the hybridization yield, despite a slight difference in the rate, however it does increase the duplex stability dramatically. The noncovalent nucleic acid–graphene oxide complex maintained the stability between 25 and 90 °C in the absence of oligonucleotide-induced desorption. The melting temperatures of duplexes with or without LNA base modification were determined due to remarkable fluorescence quenching and fast oligonucleotide adsorption with graphene oxide. The difference in melting temperatures was used to control the release of surface adsorbed nucleic acids at 70 °C. Finally, a mutation in the oligonucleotide sequence is detected by the complementary oligonucleotides on the graphene oxide surface. Due to its extraordinary physical properties, graphene oxide represents a remarkable platform for studying nucleic acid interactions and serves as a promising material for biomedical applications.
Graphene Oxide Protected Nucleic Acid Probes for Bioanalysis and Biomedicine
Chemistry - A European Journal, 2013
Graphene Oxide Protected Nucleic Acid Probes for Bioanalysis and Biomedicine Protect and survive! Single-stranded nucleic acids can absorb on graphene oxide (GO) surfaces and be effectively protected from enzymatic degradation. This property offers a novel way to stabilize nucleic acid probes when used in vivo for molecular probing and bio-medical applications such as siRNA and gene delivery. It also provides new opportunities for amplified detection of small molecules and microRNA in vitro with high sensitivity and selectivity. Nucleic Acid Probes In their Concept article on page && ff., Z. Zhu, C. J. Yang et al. give an overview of the latest developments in the use of graphene oxide in bioanalysis and biomedicine, focusing on the use of GO-protected nucleic acid probes.
Progress in miRNA Detection Using Graphene Material–Based Biosensors
Small, 2019
the potential to trigger carcinogenic pathways and various genetic changes. [13] To explore the pathogenetic mechanisms and achieve precise disease diagnoses, tremendous efforts have been exerted, among them computed tomography (CT), radiography and blood tests are considered the most effective methods in clinical applications. [14-16] However, because of bulky instrumentation, expense, complex sample preparation, and tedious operation of instruments, these methods require experienced operators, high-throughput capacity and are time consuming. In addition, these detection methods are typically accompanied by radiation and trauma during the detection process, which are harmful to the human body. Moreover, in most cases, only large tumors can be observed using these imaging techniques, and thus, most cancers are detected at a middle or advanced stage, and cancer is less likely to be diagnosed at a more treatable stage. Therefore, more researchers are focusing on detecting tumor biomarkers because it is a new type of platform that addresses early diagnosis and treatment of cancer. Biomarker selection and research plays an important role in biomedical research. MicroRNAs (miRNAs) are small (18-22 nucleotides) noncoding single-stranded RNA molecules that play key roles in cell proliferation, differentiation, and apoptosis. [17,18] In addition, studies have shown that abnormal expression of certain miRNAs is closely related to tumor occurrence, tumor staging, and tumor treatment response, [19-22] and the presence of miRNA in serum can be clearly detected. [23] All these findings suggest that miRNAs can be used as next-generation tumor markers with clinical diagnostic and prognostic significance for early diagnosis of malignant disease. [24] To truly treat miRNA as a useful biomarker in clinical practice, it is important to develop sensing technology to quantitatively measure the expression level of specific miRNAs. Currently, the widely used miRNA analysis methods, including real-time reverse transcription polymerase chain reaction, [25] quantitative real-time polymerase chain reaction (qRT-PCR), northern blotting, [26] and miRNA array technologies, [27] can meet the detection requirements to a certain extent. However, exosome destruction and RNA extraction using these traditional biotechnologies require expensive commercial kits, which contain abundant interfering RNAs. Moreover, the concentration of miRNA may not be adequate for detection, and the high cost makes many patients flinch. [28] MicroRNAs (miRNAs) are short, endogenous, noncoding RNAs that play critical roles in physiologic and pathologic processes and are vital biomarkers for several disease diagnostics and therapeutics. Therefore, rapid, low-cost, sensitive, and selective detection of miRNAs is of paramount importance and has aroused increasing attention in the field of medical research. Among the various reported miRNA sensors, devices based on graphene and its derivatives, which form functional supramolecular nanoassemblies of π-conjugated molecules, have been revealed to have great potential due to their extraordinary electrical, chemical, optical, mechanical, and structural properties. This Review critically and comprehensively summarizes the recent progress in miRNA detection based on graphene and its derivative materials, with an emphasis on i) the underlying working principles of these types of sensors, and the unique roles and advantages of graphene materials; ii) state-of-the-art protocols recently developed for high-performance miRNA sensing, including representative examples; and iii) perspectives and current challenges for graphene sensors. This Review intends to provide readers with a deep understanding of the design and future of miRNA detection devices. Graphene Biosensors Congcong Zhang obtained her Ph.D. degree in 2012, in the Department of Chemistry,
Graphene Oxide Modified Chemically Activated Graphite Electrodes for Detection of microRNA
Electroanalysis, 2017
In our study, graphene oxide (GO) modified graphite electrodes were used for sensitive and selective impedimetric detection of miRNA. After chemical activation of pencil graphite electrode (PGE) surface using covalent agents (CA), GO modification was performed at the surface of chemically activated PGE. Then, CA‐GO‐PGEs were applied for impedimetric miRNA detection. The microscopic and electrochemical characterization of CA‐GO‐PGEs was performed by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). The optimization of experimental conditions; such as GO concentration, DNA probe concentration and miRNA target concentration was performed by using EIS technique. After the hybridization occurred between miRNA‐34a RNA target and its complementary DNA probe, the hybrid was immobilized onto the surface of CA‐GO‐PGEs. Then, the impedimetric detection of miRNA‐DNA hybridization was performed by EIS. The selectivity of our assay was also tested under the opti...
Graphene oxide modified single-use electrodes and their application for voltammetric miRNA analysis
Materials science & engineering. C, Materials for biological applications, 2017
The modification of graphene oxide (GO) onto the surfaces of chemically activated pencil graphite electrodes (PGEs) was performed herein, and then these electrodes were applied for the first time on voltammetric monitoring of miRNAs. The specific recognition of miRNA-34a, which has been related to Alzheimer disease, was explored in the presence of DNA-RNA hybridization by using CA/GO/PGEs in combination with differential pulse voltammetry (DPV) technique. The characterization of CA/GO/PGE was investigated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and scanning electron microscopy (SEM). The effect of GO concentration, DNA probe concentration and miRNA-34a concentration upon to biosensor response was optimized, and accordingly, the selectivity of miRNA-34a biosensor was tested under the optimum conditions.
Electrochemical detection of microRNAs by graphene oxide modified disposable graphite electrodes
Journal of Electroanalytical Chemistry, 2018
Graphene oxide (GO) modified disposable pencil graphite electrodes (PGEs) were developed for the first time in the present study for miRNA detection by measuring the guanine oxidation signal. GO modified PGEs were used as solid phase to immobilize amino linked DNA probe and then the complementary target RNA sequence (miRNA-34a) was recognized. The electrochemical behaviour of the GO modified electrodes was investigated by using electrochemical impedance spectroscopy (EIS) and microscopic characterization was performed by scanning electron microscopy (SEM). The optimization studies and detection of hybridization between miRNA-34a target and its complementary DNA probe was performed by differential pulse voltammetry (DPV). The selectivity of our assay was tested in the presence of non-complementary miRNA sequence. A good discrimination could be achieved compared to the results obtained with the full match hybridization of probe with its RNA target miRNA-34a target against other miRNA sequences; miRNA-16 and miRNA-155.
Analytical Chemistry, 2013
RNA probes constitute an important class of functional nucleic acids (FNAs). However, because of their notorious vulnerability to enzymatic degradation, extremely careful and special protocols must be followed when dealing with RNA probes. To fully use the large number of RNA FNAs available for bioanalysis and biomedicine, it is important to explore effective methods to protect RNA probes from enzymatic digestion. In this work, we systematically demonstrate that graphene oxide (GO) can effectively protect RNA probes from enzymatic digestion. Based on this finding, we propose an effective way to design robust RNA biosensors by simply mixing RNA probes with GO for analysis of nucleic acids, proteins, and small molecules. The entire assay is sensitive, selective, rapid, and more importantly, does not require any special protocols. The ability to protect ssRNA from enzymatic digestion by GO offers an exciting new way to stabilize ssRNA, which will not only provide new opportunities to utilize the large number of currently available, yet rarely explored, RNA FNAs for bioanalysis but also offer a new solution to protect important ssRNA molecules, such as microRNA and antisense ssRNA, for a great variety of biomedical applications.
MRS Communications, 2018
Nanomaterials have been proposed as key components in biosensing, imaging, and drug-delivery since they offer distinctive advantages over conventional approaches. The unique chemical and physical properties of graphene make it possible to functionalize and develop protein transducers, therapeutic delivery vehicles, and microbial diagnostics. In this study we evaluate reduced graphene oxide (rGO) as a potential nanomaterial for quantification of microRNAs including their structural differentiation in vitro in solution and inside intact cells. Our results provide evidence for the potential use of graphene nanomaterials as a platform for developing devices that can be used for microRNA quantitation as biomarkers for clinical applications.
Sensors and Actuators A-physical, 2018
Highlights EDC-NHS surface chemistry was applied for the preparation of GO modified PGEs. PGEs were used to detect miRNA-34a which is a biomarker for Alzheimer and cancer. Step-by-step hybridization was applied to obtain a controllable process. EIS provided a new aspect to detect miRNA-34a in contrast to traditional methods. The detection of miRNA-34a in FBS was also explored with a low detection limit.
Analytical Chemistry, 2013
We report a simple and sensitive label-free immunosensor for detection of microRNAs (miRNA) based on a conducting polymer/reduced graphene oxide-modified electrode to detect miR-29b-1 and miR-141. Square wave voltammetry is used to record the redox signal. Current increases upon hybridization (signal on) from 1 fM to 1 nM of target miRNA. The limit of quantification is ca. 5 fM. The sensor exhibits high selectivity as it distinguishes mismatch. To double-check its selectivity, two specific RNA−DNA antibodies recognizing miRNA−DNA heteroduplexes, antipoly(A)−poly-(dT) and anti-S9.6, were used. The antibody complexation with the hybrid leads to a current decrease that confirms the presence of miRNA, down to a concentration of 8 fM. The antibody−hybrid complex can be then dissociated by adding miRNA−DNA hybrids in solution, causing a shift-back on the signal, i.e., an increase in the current density (signal-on). This On−Of f−On detection sequence was used as a triple verification to increase the reliability of the results.