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Highlights: [•] First aptasensor based on SAMs’ interfacial energy transfer was reported. [•] High quenching efficiency of GO and SAMs’ concentration lead to higher sensitivity. [•] SAMs is easily designed and reduces the labor for the labeling of aptamer to QDS. [•] This universal and simple platform has the potential for sensitive biosensing. [ABSTRACT FROM AUTHOR]

MiR-451a is a biomolecule tightly related to resistance development during treatment with selective estrogen-receptor modulators for breast cancer. Accurately and sensitively monitoring its level in cells is of great significance for the drug selection and the evaluation of this cancer. However, the commonly obtainable ssDNA probe cannot efficiently enter the cell to sensitively emit reliable signals by avoiding nuclease digestion. Herein, a sensing system is reported to monitor miR-451a in living cells with high specificity and sensitivity. The system consisted of reduced graphene oxide (rGO), duplex-specific nuclease (DSN), and a fluorescence-labeled ssDNA nanoprobe. The fluorescence probe on the rGO surface provided an ultra-low background and resistance to endonuclease digestion. Duplex-specific nuclease sequentially triggered the hybridization cycle and the cleavage of ssDNA nanoprobe. The signal amplification provided a detection limit of 1 fM miR-451a using the optimal conditions with a linear range from 1 fM to 100 nM. Finally, the developed method was used to determine miR-451a in breast cancer tissue. This strategy provides a promising tool for clinical diagnosis and evaluation of miR-451a related cancer. [ABSTRACT FROM AUTHOR]

We herein find that graphene oxide (GO) can enhance the fluorescence signal of conjugated polyelectrolytes (CPEs) with charge transfer characteristics under certain conditions, which is due to a relative hydrophobic environment of CPEs upon complexation with GO. [ABSTRACT FROM AUTHOR]

As an essential phosphate group hydrolase, alkaline phosphatase (ALP), whose level in serum is correlated with bone disease, liver dysfunction, and cancer, could be used as a biomarker for clinical diagnosis and biomedical studies. Hence, developing a convenient and sensitive method for ALP assay has importance in disease diagnosis, drug treatment, and prognosis assessment. In this work, using a hairpin DNA strand as the substrate, we developed an ultrasensitive and simple fluorescence method for quantitative ALP assay based on the binding difference of reduced graphene oxide (rGO) with different DNA strands coupled with λ exonuclease (λ exo) cleavage. Under the optimal conditions, the limit of detection (LOD) of ALP is estimated to be 0.01 U/L with the linear region from 0.5 U/L to 70 U/L. Furthermore, the proposed assay was used to detect ALP in complicated cell-free extracts and evaluate the inhibitory effects of two well-known inhibitors of ALP activity. Finally, the method was used to investigate the effect of natural compounds on ALP activity and five compounds with different inhibitory capability were screened. In summary, we propose that the new method for ALP assay can be applied for therapeutic drug monitoring (TDM) and high-throughput compound screening in combination with multiwell plate technology. [ABSTRACT FROM AUTHOR]

Mung bean nuclease is a single stranded specific DNA and RNA endonuclease purified from mung bean sprouts. It yields 5′-phosphate terminated mono- and oligonucleotides. The activity level of this nuclease can act as a marker to monitor the developmental process of mung bean sprouts. In order to facilitate the activity and physiological analysis of this nuclease, we have developed a biosensing assay system based on the mung bean nuclease-induced single-stranded DNA scission and the affinity difference of graphene oxide for single-stranded DNA containing different numbers of bases. This end-point measurement method can detect mung bean nuclease in a range of 2×10 −4 to 4×10 −2 with a detection limit of 1×10 −4 unit/mL. In addition, we demonstrate the utility of the assay for screening chemical antibiotics and metal ions, resulting in the identification of several inhibitors of this enzyme in vitro . Furthermore, we firstly report that inhibiting mung bean nuclease by gentamycin sulfate and kanamycin in vivo can suppress mung bean sprouts growth. In summary, this method provides an alternative tool for the biochemical analysis for mung bean nuclease and indicates the feasibility of high-throughput screening specific inhibitors of this nuclease in vitro and in vivo . [ABSTRACT FROM AUTHOR]

Due to the significant role of formamidopyrimidine DNA glycosylase (Fpg) in physiological processes and DNA oxidative damage-related diseases, it is essential to establish sensitive methods for monitoring the Fpg activity in vitro and in vivo so as to illustrate its concrete role in these events. In this work, a sensitive, simple and reliable fluorescence assay was developed by taking the advantages of DNAzyme assisted cascade signal amplification and ultra-high fluorescence quenching efficiency of reduced graphene oxide (rGO). This detection system consisted of DNAzyme, rGO and fluorescence probe allows the activity of Fpg to be detected in a linear range from 0 to 80 U/mL with a detection limit of 0.66 U/mL. With the help of this method, 11 natural compounds were screened, and 7 compounds were identified as activators of Fpg. More importantly, the developed assay was used to monitor the activity of Fpg through fluorescence imaging in living Escherichia coli for the first time. The imaging results visually demonstrated the dynamic activation effect of natural compound Ginsenoside Re on the Fpg of Escherichia coli. In summary, these results indicated that this DNAzyme and rGO based fluorescence assay provides a potent strategy for Fpg quantitative assay in vitro and real-time monitoring in living bacteria, which holds great potential for applying on biological study and Fpg-targeted drug screening. Image 1 • A sensitive, simple and reliable fluorescence assay was designed by taking advantage of DNAzyme and rGO. • The developed assay was applied to screen the Fpg-targeted regulators from natural compounds. • Thisfluorescence assay can realize real-time monitoring of Fpg activity in living E. coli.. • The Fpg-activation effect of natural compound Ginsenoside Re in E. coli was revealed by proposed imaging strategy. [ABSTRACT FROM AUTHOR]