ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Jiaxin Su,Jinya Du,Rujiao Ge,Chenyang Sun,Yuchun Qiao,Wei Wei,Xuejiao Pang,Yufan Zhang,Huiting Lu,Haifeng Dong
DNAzyme shows great promise in designing a highly sensitive and specific sensing platform; however, the low cellular uptake efficiency, instability, and especially the insufficient cofactor supply inhibit the intracellular molecule sensor applications. Herein, we demonstrate a novel type of DNAzyme-based self-driven intracellular sensor for microRNA (miRNA) detection in living cells. The sensor consists of a metal–organic framework [zeolite imidazole framework (ZIF-8)] core loaded with a shell consisting of a rationally designed DNAzyme, where the substrate strand is modified with FAM and BHQ-1 nearby both the sides of the restriction site, respectively, while the enzyme strand consists of two separate strands with a complementary fragment to the substrate strand and the targeting miRNA, respectively. The ZIF-8 nanoparticles enable the efficient delivery of DNAzyme into the cell and protect the DNAzyme from degradation. The pH-responsive ZIF-8 degradation is accompanied with the release of the DNAzyme and Zn2+ cofactors, and the intracellular target miRNAs recognize and activate the DNAzyme driven by the Zn2+ cofactors to cleave the substrate strand, resulting in the release of the FAM-labeled shorter product strand and increased fluorescence for miRNA detection. The self-driven approach can be generally applied to various miRNAs’ detection through DNAzyme engineering.
研究领域:
生物化学
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分子生物学
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纳米技术
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生物医学工程
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分析化学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Wanqing Yu,Fangjing Mo,Jiang Guo,Yuqin Yang,Yushuang Jin,Yingzi Fu
Seeking and constructing superior photoactive materials have the potential to improve the performance of photoelectrochemical (PEC) biosensors. In this work, we proposed a novel mimosa-like ternary inorganic composite with a significantly enhanced light-harvesting ability and photogenerated carrier separation rate. This ternary photoactive material was obtained via electrodeposition of gold nanoparticles (Au) on the surface of transition metal sulfide composite of CdS and NiS (CdS-NiS/Au). The experimental results showed that the high initial photocurrent was acquired on CdS-NiS/Au (68-fold higher than that of individual CdS) with the synergistic effect of p-n heterojunction, Schottky junction, and the eminent optical properties of gold nanoparticles. Meanwhile, using silver nanoclusters prepared by link DNA protection as an effective quencher, integrating the duplex-specific nuclease-assisted rolling circle amplification strategy, a “Signal ON” PEC biosensor was fabricated for the detection of microRNA 21 (miRNA 21). With the release of the quencher, the recovered photocurrent is able to achieve determination of miRNA 21 within the range from 10 aM to 1 pM with a detection limit down to 4.6 aM (3σ). Importantly, this work not only provides a superb idea for designing ternary inorganic heteromaterials with exceptional photoactive ability but also allows the detection of other biomarkers by selecting appropriate recognition units.
研究领域:
生物传感技术
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纳米材料科学
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分子诊断学
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Xing Huang,Zihao Li,Yakun Shi,Yanfei Zhang,Taorong Shen,Meng Chen,Zhan Huang,Yanli Tong,Si-Yang Liu,Jianhe Guo,Xiaoyong Zou,Zong Dai
Autocatalytic biocircuit are powerful tools for analysing intracellular biomarkers, but these tools are constrained by limitations in amplification capacity and intracellular delivery efficiency. In this work, we developed a DNAzyme-based dual-feedback autocatalytic exponential amplification biocircuit sustained by a honeycomb MnO 2 nanosponge (EDA 2 @hMNS) for live-cell imaging of intracellular low-abundance microRNAs (miRNA). The EDA 2 biocircuit comprises a blocked DNAzyme ( b -DNAzyme), a Fuel strand and a Substrate strand. In the EDA 2 biocircuit, target miRNAs are recycled and feedback for rounds of DNAzymatic amplification, and the DNAzymatic reactions continuously generate target miRNA analogues for dual-feedback to achieve multiple parallel cascade DNAzymatic reactions that improve amplification capacity substantially. In addition, the hMNS ensures high loading and delivery efficiency of biocircuit probes into living cells and also provides sufficient Mn 2+ DNAzyme cofactor from in situ decomposition by intracellular glutathione (GSH). The EDA 2 @hMNS realized a detection limit of 17 pM, which is 288-fold lower than the b -DNAzyme lacking the DNAzymatic amplification. These results demonstrate the great promise for this critical tool in analysing low-abundance biomarkers and cancer diagnostics.
研究领域:
分子生物学
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生物化学
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癌症诊断
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Yong Wang,Dandan Wang,Guohua Qi,Ping Hu,Erkang Wang,Yongdong Jin
As one of the most widely distributed microRNAs, microRNA-21 (miRNA-21) significantly regulates target genes’ expression levels and participates in many cellular and intercellular activities, and its abnormal expression is always related to some diseases, especially cancer. Hence, detecting miRNA-21, as a biomarker, at the single-cell level helps us to reveal cell heterogeneity and expression level variation during the state change of cells. In this study, we constructed a gold nanoparticles nanomembrane (AuNPs-NM)-modified plasmonic glass nanopipette (P-nanopipette) surface-enhanced Raman scattering (SERS) sensing platform to sensitively detect content variation of the intracellular miRNA-21 during the electrostimulus (ES)-induced apoptosis process. The cytoplasm-located miRNA-21 was first extracted by using the extraction DNA (HP1)-modified P-nanopipette through a hybridization chain reaction (HCR). The nanopipette was then incubated with a labeling DNA (HP2) and reporter 4-MBA-modified Raman tag. The Raman signal (collected from the tip area near the orifice within 1 μm) showed a good response to the content variation of intracellular miRNA-21 under ES, and the proposed single-cell SERS detection platform provides a simple way to study intracellular substance change and evaluate cancer treatment outcomes.
研究领域:
生物技术
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纳米技术
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癌症研究
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分子生物学
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分析化学
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Zhengfang Liang,Yunzhan Xie,Hongbing Zhou,Zilu Zhang,Yuanxun Gong,Jiayi Zhang,Yue Huang,Qianli Tang,Kai Zhang,Xianjiu Liao
MicroRNA-21 (miR-21) is a critical biomarker involved in various pathological processes, yet its low abundance in biological fluids poses challenges for accurate detection. Herein, we present a highly sensitive electrochemiluminescence (ECL) biosensor for miR-21 detection. The system integrates a multi-stage amplification cascade that begins with toehold-mediated strand displacement (TMSD) and continues through T7 RNA polymerase transcription and CRISPR/Cas13a-mediated collateral cleavage. The sensing interface was constructed by immobilizing ferrocene-labeled DNA probes onto a conductive AuNPs/Ti₃C₂Tx/Ru(II)-PEI nanohybrid-modified electrode. In the absence of miR-21, ferrocene effectively quenched the ECL signal. Upon target recognition, TMSD activates a split T7 promoter/template system, triggering RNA transcription. The resulting single-stranded RNAs activate Cas13a, which cleaves DNA probes and restores the ECL emission. Under optimized conditions, the biosensor exhibited a wide dynamic range from 10 aM to 500 fM and a detection limit as low as 5.23 aM. It also demonstrated excellent specificity, repeatability (RSD = 4.3 %), and recovery (99.93–106.20 %) in spiked serum samples. This work provides a versatile and modular ECL sensing platform for ultrasensitive miRNA detection, offering promising applications in clinical diagnostics and precision medicine.
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DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2018
·
IF 6.042
Ting Hou,Ningning Xu,Wenxiao Wang,Lei Ge,Feng Li
In conventional photoelectrochemical (PEC) analysis, photoactive materials are usually immobilized on electrode surfaces, and such immobilization procedures are tedious and time-consuming, and it is also difficult to prepare electrodes with good reproducibility. To circumvent such limitations, we propose here a truly immobilization-free diffusivity-mediated PEC bionsensing strategy for microRNA assay, using methylene blue (MB) in solution as the photoactive probe, and nonmodified indium tin oxide (ITO) glass as the working electrode. The hybridization between the target microRNA and the MB-labeled single-stranded DNA probe (MB-DNA) triggers the digestion of MB-DNA by T7 exonuclease (T7 Exo), thus to generate MB-labeled mononucleotide, and then the released target microRNA initiates the subsequent cycling processes and generates a large amount of MB-labeled mononucleotides. Due to the diffusivity difference between MB-DNAs and MB-labeled mononucleotides, significantly increased photocurrent signal is observed for MB-labeled mononucleotides as compared to that of MB-DNAs. Therefore, via this “signal-on” mode and the T7 Exo facilitated signal amplification, a facile and highly sensitive immobilization-free PEC microRNA assay is readily realized, with a detection limit down to 27 aM. Moreover, this strategy exhibits excellent specificity and is successfully applied in detecting microRNA spiked in serum samples. Since all the reactions take place in homogeneous solutions and no electrode modification is needed, this PEC biosensing strategy exhibits the advantages of simplicity, rapidness, and good reproducibility. More significantly, it provides a novel concept to design truly immobilization-free PEC biosensing systems, and shows potential to be applied in bioanalysis and biochemical research.
研究领域:
生物分析
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分子生物学
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DOI
高影响因子
Nanoscale
·
2018
·Vol. 10
·期 15
·7067-7076
·
IF 7.233
Jiantao Yu,Sihui He,Chen Shao,Haoran Zhao,Jing Li,Leilei Tian
The design of a nano-system for the detection of intracellular microRNAs is challenging as it must fulfill complex requirements, i.e., it must have a high sensitivity to determine the dynamic expression level, a good reliability for multiplex and simultaneous detection, and a satisfactory biostability to work in biological environments. Instead of employing a commonly used physisorption or a full-conjugation strategy, here, a GO-DNA nano-system was developed under graft/base-pairing construction. The common anchor sequence was chemically grafted to GO to base-pair with various microRNA probes; and the hybridization with miRNAs drives the dyes on the probes to leave away from GO, resulting in “turned-on” fluorescence. This strategy not only simplifies the synthesis but also efficiently balances the loading yields of different probes. Moreover, the conjugation yield of GO with a base-paired hybrid has been improved by more than two-fold compared to that of the conjugation with a single strand. We demonstrated that base-paired DNA probes could be efficiently delivered into cells along with GO and are properly stabilized by the conjugated anchor sequence. The resultant GO-DNA nano-system exhibited high stability in a complex biological environment and good resistance to nucleases, and was able to accurately discriminate various miRNAs without cross-reaction. With all of these positive features, the GO-DNA nano-system can simultaneously detect three miRNAs and monitor their dynamic expression levels.
研究领域:
分子生物学
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纳米医学
·
生物化学
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DOI
PubMed
高影响因子
BIOELECTROCHEMISTRY
·
2025
·
IF 4.5
Jinshan Liu,Songyang Miao,Niewei You,Jinghan Li,Siao Li,Xun Chen,Jie Wang,Mu Yang,Kejian Ding
Early cancer diagnosis is critically enhanced by detecting microRNA (miRNA) biomarkers, such as miRNA-155, whose overexpression correlates with cancer progression. This work presents a highly sensitive photoelectrochemical (PEC) biosensor for miRNA-155 detection leveraging a modified carbon nitride (m-CN) substrate modified by ReS₂ (ReS₂/m-CN), which enhanced electron transfer efficiency and provided abundant sites. The resulting biosensor exhibits exceptional performance with a broad linear detection range from 10 −16 M to 10 −7 M, an ultralow detection limit, high selectivity, and outstanding stability. This work demonstrates the significant potential of the ReS 2 @m-CN-based PEC biosensor for sensitive, cost-effective early cancer diagnostics.
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产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2024
·
IF 8.3
Xiaojian Fan,Youqiang Xing,Ze Wu,Bingjue Li,Peng Huang,Lei Liu
Sensitive and accurate miRNA detection is important in cancer diagnosis but remains challenging owing to the essential features of miRNAs, such as their small size, high homology, and low abundance. This work proposes a novel electrochemical (EC)-enhanced quantum sensor achieving quantitative detection of miRNA-155 with simultaneous EC sensing. Specifically, fluorescent nanodiamonds/MXene nanocomposites were synthesized and modified with dual-mode signal labels, enabling miRNA-155 concentration measurement via T1 relaxation time of nitrogen-vacancy (NV) centers and EC signals. Quantum sensing was enhanced via external voltage during the EC process, which modulated the negatively charged state of the NV centers, thereby improving the sensitivity and accuracy of miRNA-155 detection. EC sensing improved the accuracy and reliability of miRNA-155 detection while enhancing quantum sensing. The limit of detection (LOD) of the EC-enhanced quantum biosensor reached 10.0 aM, nearly 106 and 10 times lower than the reported LODs of a quantum sensor using bulk diamond and fluorescent sensors, respectively. The LOD of EC sensing was 2.6 aM, aligning with previous reports. The findings of the study indicated that quantum sensing combined with EC sensing can achieve ultrasensitive miRNA-155 detection with high accuracy and reliability, providing an advanced approach for early cancer diagnosis.
研究领域:
生物医学工程
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分子生物学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
ACS Omega
·
2024
·
IF 3.7
Ling Yang,Chunxiang Che,Mengfan Guo,Chunwei Fan,Lina Sun,Shanshan Chen
MicroRNAs (miRNAs), a class of small molecules with important regulatory functions, have been widely used in biosensing as biomarkers for the early diagnosis of various diseases (such as osteoporosis). Therefore, it is necessary to develop a miRNA detection platform with high sensitivity and specificity. Herein, an upconversion biosensor based on fluorescent resonant energy transfer (FRET) for the detection of miRNAs was developed. Gold nanoparticles (AuNPs, fluorescence acceptors) were conjugated with aptamers, while upconversion nanoparticles (UCNPs, fluorescence donors) were functionalized with the corresponding complementary DNA (cDNA). The upconversion luminescence (UCL) is quenched vis FRET due to the good overlap between UCNP fluorescence emission and AuNP absorption. In the presence of the target miRNA, the aptamers preferentially bind to the miRNA, thereby dissociating the UCNP-cDNA from the AuNP-aptamers, resulting in the recovery of the UCL. The UCL recovery efficiency increased in a miRNA concentration-dependent manner, which supported the principle of miRNA quantification. The UCNP-based FRET biosensor exhibited sensitive detection of miRNA-21 (as a model analyte) with low limit of detection of 5.91 pM. In addition, depending on the near-infrared excitation and a simple complementary binding reaction during the detection process, the designed biosensor not only effectively eliminated the autofluorescence but also reduced the detection time.
研究领域:
生物化学
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分子生物学
·
纳米医学
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产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2024
·
IF 8
Yuqin Yang,Gui Zhang,Li Song,Guangrong Kuang,Yilan Ding,Shuchun Bu,Yaqin Chai,Yingzi Fu
Carbon dots (CDs) with excellent optical performance have attracted tremendous interest for the construction of electrochemiluminescence (ECL) biosensors. However, the intrinsic self-aggregation behavior of CDs brings a serious issue of aggregation-caused quenching (ACQ), which can quench the ECL emission. In this work, different surfactants (non-ionic, anionic and cationic) were used to modulate the preparation of functionalized CDs (CDs-based nanocomposites). Their ECL performances were significantly enhanced in potassium persulfate solution. In particular, the highest ECL intensity obtained from the Triton X-100 functionalized CDs (T-CDs) was 16.8-fold than that of CDs as ACQ was suppressed effectively. Therefore, utilizing T-CDs as an ECL luminophore, integrating with the catalytic hairpin assembly and strand displacement reaction signal amplification strategies, a "Signal on" ECL biosensor was constructed for microRNA-21 (miRNA-21) monitoring. The proposed biosensor shows a dynamic range from10 fmol·L −1 to 10 nmol·L −1 at a low detection limit of 2.9 fmol·L −1 (S/N = 3). Meanwhile, it exhibits good results for analyzing miRNA-21 in healthy human samples and the expression in MCF-7 and HeLa cells. This work not only expands the method for suppressing ACQ of CDs, but also shows great potential for the analysis of other biomarkers by replacing the corresponding DNA sequences , so as to realize the early diagnosis.
研究领域:
生物传感技术
·
分子诊断学
·
纳米生物学
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Xuemei Yin,Qindong Guo,Xingqi Ji,Xiaoqian Li,Hao Xue,Qian Xin,Jiawei Zhang,Zhuocheng Yan,Aimin Song
MicroRNAs have started being used as an effective marker in early diagnosis and preoperative monitoring of cancers in recent years. Traditional microRNA testing technology, such as quantitative reverse transcription polymerase chain reaction (qRT-PCR) and various fluorescent and colorimetric assays, often suffer from complicated operations and limited sensitivity. They also produce significant electronic waste posing threats to human health and environment. Here, we propose a degradable biosensor using indium-gallium-zinc oxide (IGZO) thin-film transistor (TFT) arrays for rapid and highly sensitive detection of microRNAs from glioma exosome extracts (g-miRNAs). The IGZO transistor arrays fabricated on nanofibrillated cellulose paper exhibit high electronic performance with a current on/off ratio >1.5 × 10 6 , a mobility of 9.6 cm 2 V −1 ·s −1 , a subthreshold swing <0.5 V·dec −1 , and excellent bias stress stability. Specific DNA probes in the IGZO channel bind selectively with g-miRNA targets to form DNA-RNA double strands, offering high specificity even when coexisting with high concentrations of nonspecific microRNAs. The inherent negative charges in DNA and g-miRNA molecules sensitively modulate the IGZO channel conductivity, leading to a positive shift of threshold voltage and decrease of source-drain current. These changes are linearly correlated with microRNA concentrations from 1 fM to 100 pM, with a detection limitation of 350 aM. Furthermore, the paper-based IGZO transistor array nearly completely dissolves in a NaOH solution after 300 min. The proposed approach combines easy-to-operate, point-of-care microRNA testing with lightweight, low-cost, biocompatible, and degradable devices, showing great promise for early diagnosis of glioma and most likely also other tumors.
研究领域:
生物医学工程
·
材料科学
·
肿瘤学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）