Nanoscale
·
2019
·Vol. 11
·期 9
·4044-4052
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IF 6.97
Lijun Geng,Xudong Yu,Yajuan Li,Yanqiu Wang,Yongquan Wu,Jujie Ren,Fengfeng Xue,Tao Yi
Biomolecule-based hydrogels have potential use in a wide range of applications such as controlled drug release, tissue engineering, and biofabrication. Herein, driven by specific interactions between ds-DNA (double-stranded DNA) and Zn2+ based metal-complexes, we report that the use of DNA as cross-linkers can enhance interactions between self-assembling Zn2+ complexes containing terpyridine and sugar groups in the generation of bioinspired hydrogels from solutions or suspensions. The gelation process is fast and straightforward without tedious steps and happens at room temperature. Such a hydrogelation process of different Zn2+ complexes endows the visualized and selective DNA analogue discrimination. Several experiments suggest that the strong intercalation binding of Zn2+ complexes with ds-DNA results in the unzipping of ds-DNA into ss-DNA (single-stranded DNA), which further behave as linkers to enhance the intermolecular interactions of self-assembling Zn2+ complex molecules via coordination interactions. This work demonstrates an efficient and universal strategy to prepare hydrogels based on biomolecular recognition. Moreover, the DNA responsive behaviors of Zn2+ complexes are further compared with that of solutions and cells.
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APPLIED SURFACE SCIENCE
·
2019
·
IF 5.155
Yujie Qiang,Shengtao Zhang,Liping Wang
Deoxyribonucleic acid (DNA) was investigated for suppressing copper corrosion in 0.5 M H 2 SO 4 through electrochemical measurement, microscopic surface observation, X-ray photoelectron spectroscopy (XPS) analysis and molecular dynamics (MD) simulation. Results indicate that DNA acts as an effective cathodic-type inhibitor that mainly suppresses cathodic reaction by forming anchored DNA-adsorption film. The film shows active blocking effect, obeys the Langmuir adsorption model and involves physisorption and chemisorption . The MD simulation gives the new insights at molecular level and indicates the formation of an interesting double barrier of DNA molecule on the Cu (111) surface due to its unique twist structure.
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ANALYTICA CHIMICA ACTA
·
2018
·
IF 5.123
Mengke Wang,Zihan Lin,Qing Liu,Shan Jiang,Hua Liu,Xingguang Su
A novel fluorescent biosensor for protein kinase activity (PKA) detection was designed by applying double-strands DNA-hosted copper nanoclusters (dsDNA-CuNCs) and graphene oxide (GO). One DNA strand of the dsDNA consisted of two domains, one domain can hybridize with another complementary DNA strand to stabilize the fluorescent CuNCs and another domain was adenosine 5′-triphosphate (ATP) aptamer. ATP aptamer of the dsDNA-CuNCs would be spontaneously absorbed onto the GO surface through π-π stacking interactions. Thus GO can efficiently quench the fluorescence (FL) of dsDNA-CuNCs through fluorescence resonance energy transfer (FRET). In the present of ATP, ATP specifically combined with ATP aptamer to form ATP-ATP aptamer binding complexes, which had much less affinity to GO, resulting in the fluorescence recovery of the system. Nevertheless, in the presence of PKA, ATP could be translated into ADP and ADP could not combine with ATP aptamer resulting in the fluorescence quenching of dsDNA-CuNCs again. According to the change of the fluorescence signal, PKA activity could be successfully monitored in the range of 0.1–5.0 U mL −1 with a detection limit (LOD) of 0.039 U mL −1 . Besides, the inhibitory effect of H-89 on PKA activity was studied. The sensor was performed for PKA activity detection in cell lysates with satisfactory results.
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ELECTROPHORESIS
·
2018
·Vol. 40
·期 3
·425-430
·
IF 2.569
Yating Zhang,Yan Zhang,Luqi Zhu,Pingang He,Qingjiang Wang
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
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ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Xudong Wang,Jun Dai,Xuehong Min,Zhihua Yu,Yong Cheng,Kaixun Huang,Juliang Yang,Xiaoqing Yi,Xiaoding Lou,Fan Xia
Detection of an ultralow concentration of mRNA is important in the prognosis of gene-related diseases. In this study, a DNA-conjugated amphiphilic aggregation-induced emission probe (TPE-R-DNA) was synthesized for cancer tissue imaging and prognosis analysis based on an exonuclease III-aided target recycling technique. TPE-R-DNA comprise two components: a hydrophobic component that serves as the “turn-on” long wavelength fluorescence imaging agent (TPE-R-N3); and a hydrophilic single DNA strand (Alk-DNA) which acts as specific recognition part for target mRNA. In the absence of target mRNA, TPE-R-DNA had almost no fluorescence because of its high water solubility. Conversely, the TPE-R-DNA was digested by exonuclease III (Exo III) in the presence of MnSOD mRNA to release the hydrophobic fluorogens (TPE-R-AT). Subsequently, TPE-R-AT formed aggregates, and therefore, fluorescence signal was distinctly observed. For the first time, the structure of the hydrolysis product (TPE-R-AT), containing two bases A and T, was proved by the mass spectrum (MS) and high-performance liquid chromatography (HPLC). Moreover, the detection limit toward mRNA could be achieved in as low as 0.6 pM. Furthermore, the fluorescent signal can be used to confirm the MnSOD mRNA expression level in cancer tissue. The MnSOD mRNA expression in renal cancer was lower than in renal cancer adjacent tissue. In particular, the expression level was analyzed to predict prognosis of cancer patients. Our results demonstrate that a shorter survival time was evident among patients in lower MnSOD mRNA expression. Thereby, it indicates great potential for the development of an ultrasensitive biosensing platform for the application in disease prognosis.
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TALANTA
·
2018
·
IF 4.244
Junqing He,Junyan Wang,Nan Zhang,Luyao Shen,Linlin Wang,Xiao Xiao,Yan Wang,Tao Bing,Xiangjun Liu,Songqing Li,Dihua Shangguan
Ovarian cancer is regarded as the most lethal gynecologic malignancy with poor prognosis and high mortality rate. Drug-resistance was thought to be the main obstacle to improving overall survival rate of ovarian cancer. New ligands for drug-resistant ovarian cancer cells have potential for the development of diagnosis and therapy of ovarian cancer. In present work, we reported two aptamers , HF3–58 and HA5–68 generated by cell-SELEX, against a paclitaxel-resistant ovarian cancer cell line (A2780T). Both two aptamers exhibited high selectivity and strong affinity to target cells with low nanomolar dissociation constants . The binding of aptamers to target cells was independent of divalent ions, and was tolerant of incubation temperature and nucleases in serum. Molecular targets of the two aptamers were preliminarily demonstrated to be two different glycoproteins on cell surface of A2780T cells. Owing to the structure stability and high resistance to nuclease, these two aptamers had good performance in the detection of drug-resistant ovarian cancer cells in human serum.
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Polymers
·
2018
·Vol. 10
·期 10
·1080
·
IF 2.935
Ju-Hui Zhang,Hui-Zhen Yang,Ji Zhang,Yan-Hong Liu,Xi He,Ya-Ping Xiao,Xiao-Qi Yu
The linking and modification of low molecular weight cationic polymers (oligomers) has become an attracted strategy to construct non-viral gene carriers with good transfection efficiency and much reduced cytotoxicity. In this study, PEI 600 Da was linked by biodegradable bridges containing rigid aromatic rings. The introduction of aromatic rings enhanced the DNA-binding ability of the target polymers and also improved the stability of the formed polymer/DNA complexes. The biodegradable property and resulted DNA release were verified by enzyme stimulated gel electrophoresis experiment. These materials have lower molecular weights compared to PEI 25 kDa, but exhibited higher transfection efficiency, especially in the presence of serum. Flow cytometry and confocal laser scanning microscopy results indicate that the polymers with aromatic rings could induce higher cellular uptake. This strategy for the construction of non-viral gene vectors may be applied as an efficient and promising method for gene delivery.Keywords:non-viral vectors;gene delivery;polyethylenimine;structure–activity relationship
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Analytical Methods
·
2018
·Vol. 10
·期 32
·3933-3938
·
IF 2.073
Yuping Han,Lin Shen,Zhen Li,Zhihong Liu
Upconversion nanoparticles (UCNPs) have attracted increasing interest owing to their excellent properties in chem/bio sensing. And DNA is the most frequently used biomolecule to functionalize the nanoparticles. However, there is still an urgent need for an effective and convenient strategy to assemble DNA strands on UCNPs. Herein, we utilized polydopamine (PDA) to coat hydrophobic UCNPs by a water-in-oil microemulsion method, which were then assembled with DNA through hydrogen bonding and π–π stacking interactions between the nucleotides of DNA and the aromatic groups of the PDA shell on the surface of the UCNPs. This strategy exhibited several merits such as decreasing the random coordination between phosphates of the DNA backbone and lanthanide ions of the UCNP surface and retaining the intrinsic DNA biorecognition functions for targets. A proof-of-concept biosensor for carcinoembryonic antigen (CEA) based on luminescence resonance energy transfer (LRET) was constructed using CEA aptamer-stabilized silver nanoparticles (Apt-AgNPs) as the energy acceptor. Due to the strong affinity between CEA and its aptamer, the upconversion luminescence was recovered in a [CEA]-dependent manner. This facile assembly strategy can be extended to various targets and sensors by altering the aptamer.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Shaohua Ma,Yufang Hu,Qingqing Zhang,Zhiyong Guo,Sui Wang,Qingpeng Shen,Chunbo Liu,Zhihua Liu
We present a novel and simple versatile electrochemical platform for investigating some DNA-related enzymes activity based on Au(III)-coordinated adenine complex (noted as Adenine/Au complex). The synthesized Adenine/Au complex, which possesses good biocompatibility, active electrochemical performance and strong bio-affinity, enables the construction of electrochemical biosensor and further concurrently being exploited for probing DNA-related enzyme activity. The significant strategy is based on three favorable properties: the dazzling polymeric ability of terminal deoxynucleotidyl transferase (TdT); the strong adsorption interaction between TdT-yielded T-rich DNA with Adenine/Au complex; the unique electro-catalytic property of Adenine/Au complex. Here, the change in electrochemical response depends on the concentration of TdT or alkaline phosphatase (ALP), so the same novel electrochemical sensor for detecting TdT or ALP can be established with a lower detection limit. Nay, the two developed biosensors display high selectivity, good reproducibility, stability and applicability in inhibitor or real sample investigations. These results prove that the presented Adenine/Au complex gives new insight in multiple-functionally biosensing, further promoting the potential application of Adenine/Au complex-relevant nano-materials in clinical diagnosis.
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SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Chen Li,Xuejuan Chen,Zhe Zhang,Jilin Tang,Bailin Zhang
Aptamer-based microcantilever (ML) array sensor is a recently development method for the analysis of various molecules with free-label, high sensitivity and selectivity. Since its signal-transducing principle is the changes of surface stress or resonance frequency caused by molecules binding on the surface, there have difficulties in determining some low molecular weight substances. Herein, a novel detecting dopamine (DA) method based on aptamers functionalized-ML array using gold nanoparticle (NP)-DNA conjugates as the signal-amplifying strategy was developed. In this method, thiolated DA aptamers were self-assembled on the surface of ML array and then Au NP-DNA conjugates which contained a stretch of complementary sequences for DA aptamers were hybridized to DA aptamers. When DA sample interacted with its aptamer and replaced the Au NP-DNA conjugates, the surface stress of ML array was enhanced about fifteen times and resulted in much more deflection. The deflection was linearly proportional to DA concentration in the range from 0.5 to 4 μM with a lowest limit of detection 77.3 nM ( R 2 = 0.993). Some structural and functional DA analogues did not cause interference for the detection of DA. These results proved that the functionalized-ML array sensor based on Au NP-DNA conjugates could be used as a promising tool to detect low-weight molecules with high sensitivity and selectivity.
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Polymers
·
2018
·Vol. 10
·期 10
·1060
·
IF 2.935
Miao-Miao Xun,Zheng Huang,Ya-Ping Xiao,Yan-Hong Liu,Ji Zhang,Ju-Hui Zhang,Xiao-Qi Yu
Rapid enzymatic degradation and fragmentation during DNA administration can result in limited gene expression, and consequently, poor efficacy. It is necessary to use novel vectors for DNA delivery. Herein, we aimed to design useful carriers for enhancing transfection efficiency (TE). These lipopolymers were prepared through Michael addition reactions from low-molecular-weight (LMW) polyethyleneimine (PEI) and linkers with three kinds of steroids. Agarose gel electrophoresis assay results displayed that the three lipopolymers could condense plasmid DNA well, and the formed polyplexes had appropriate sizes around 200–300 nm, and zeta potentials of about +25–40 mV. The results of in vitro experiments using HeLa, HEK293, and MCF-7 cells showed that these lipopolymers present higher TE than 25-kDa PEI, both in the absence and presence of 10% serum. Flow cytometry and confocal microscopy studies also demonstrated that these lipopolymer/DNA complexes present higher cellular uptake and intracellular distribution. The measurement of critical micelle concentration (CMC) revealed that these lipopolymers could form micelles, which are suited for drug delivery. All results suggest that the three materials may serve as hopeful candidates for gene and drug delivery in future in vivo applications.Keywords:gene vectors;Michael addition;lipopolymers;steroids
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BIOCHEMICAL ENGINEERING JOURNAL
·
2018
·
IF 3.226
Ye Yang,Siming Wang,Zixin Zhou,Ruiqi Zhang,Hao Shen,Jiayi Song,Ping Su,Yi Yang
The development of novel approaches for preparing the immobilized enzyme reactors with improved reusability and catalytic performance has attracted tremendous interest because of the extensive applications and the high cost of free enzymes. Herein, we describe a mild and versatile method for the preparation of immobilized enzyme reactors by anchoring trypsin on polydopamine (PDA) functionalized magnetic nanoparticles (MNPs) through DNA directed immobilization (DDI). This reactor exhibited prominent reusability and enhanced catalytic efficiency. The specificity constant (k cat /K m , where k cat is the catalytic rate constant) of the reactor was 1.82 s −1 μM −1 , which was approximately 4.7-fold higher than free trypsin. Notably, the immobilized trypsin reactors maintained more than 55% of their initial activity after 70 cycles (in pH 9.0 PBS, 37 °C). The improved activity and reusability were dually benefited from the rigidity and stability of DNA linkers as well as the excellent biocompatibility and cushioning effects of PDA. Given the excellent reusability and high catalytic efficiency of this immobilized trypsin reactors, we believe that this study paves the way for preparing low-cost enzyme reactors for a wide range of applications in biocatalysis, biological diagnosis, and biotechnology.
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