iScience
·
2023
·
IF 5.8
Huo Xu,Fenglin Yang,Danlong Chen,Weilin Ye,Guohui Xue,Lee Jia
MicroRNAs (miRNAs) play an important biomarker in various biological processes, especially cancer related, yet economic, simple, sensitive and specific methods for miRNA determination are still challenging. In this study, we have developed stepwise-strand displacement amplification (S-SDA)-based colorimetric sensing platform for let-7a miRNA detection in clinical serum samples. Our results demonstrated that the developed S-SDA-based method shows high sensitivity with a detection limit of 63.2 pM and a naked eye detection limit of 0.1 nM. Moreover, the S-SDA amplifier is able to discriminate target miRNAs from their mutants with high accuracy and specificity. With its high sensitivity and selectivity, this method successfully identified healthy individuals from patients with colon cancer by detecting let-7a miRNAs in serum. We believe the colorimetric analysis method will provide a new paradigm for the detection of miRNA with different abundance and show great potential for clinical application in biomedical analysis and early clinical diagnosis.
研究领域:
生物化学
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分子生物学
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临床诊断
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DOI
PubMed
高影响因子
Synthetic and Systems Biotechnology
·
2025
·
IF 4.4
Po Li,Xueying Lei,Xiaoying Niu,Wen Tian,Zhehuang Li,Songcheng Yu,Peng Zhang
MicroRNA (miRNA) is promising candidate for non-invasive diagnostic biomarker. Conventional CRISPR/Cas12a-based miRNA detection systems are constrained by reliance on reverse transcription, nucleic acid pre-amplification and costly fluorescently labeled reporters which introduce chemical modification complexity and background noise. To address these limitations, we herein developed a multifunctional switch that integrated target recognition, CRISPR/Cas12a system activation, intrinsic fluorescence signaling, and autonomous signal amplification within a single molecular architecture. As a proof of concept, this switch enabled a label-free CRISPR/Cas12a biosensing for miR-21 detection with a detection limit of 4.8 nM and robust performance in accuracy, precision, and selectivity. This proposed label-free CRISPR/Cas12a platform could be applied for real sample and is a promising candidate for point-of-care miRNA detection.
研究领域:
生物传感技术
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分子诊断
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CRISPR 基因检测
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核酸扩增
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即时检测
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2020
·Vol. 187
·期 2
·1-9
·
IF 6.232
Xiang Ling,Zhang Feng,Feng Jian,Chen Chunyan,Cai Changqun
A method is described for the simultaneous determination of hepatocellular carcinoma-associated microRNA-122 and microRNA-199a/b-3p. This probe consists of two kinds of nanomaterials. The first comprises CdTe@CdS core-shell quantum dots which, on excitation at 375 nm give two emissions, with peak wavelengths at 543 (g-QDs) and at 627 nm (r-QDs). The second comprises gold nanoparticles acting as a quencher. In the absence of the target, g-QD-N1 and r-QD-N2 are stable due to the fluorescence stability. With the addition of microRNA-122 and microRNA-199a/b-3p, g-QD-N1 and r-QD-N2 are conjugated to the surface of AuNP-S1/S2 through base complementary pairing. As a result, fluorescence resonance energy transfer (FRET) occurs, resulting in a decrease at 550 nm and 635 nm respectively, which can realize the simultaneous detection of two different microRNAs. Detection is achieved within 50 min. The detection limits (3σ/k) are 0.2 nM for microRNA-122 and 0.5 nM for microRNA-199a/b-3p. The clinical applicability of the assay was demonstrated by detecting microRNAs in human serum and different cell lysates.
Schematic for the simultaneous determination of microRNA-122 and microRNA-199a/b-3p by FRET.
研究领域:
生物化学
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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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产品关联
DOI
PubMed
高影响因子
Virulence
·
2025
·
IF 5.4
Zijing Liang,Shuoyue Wang,Xinchi Zhu,Jiale Ma,Huochun Yao,Zongfu Wu
Streptococcus suis is a Gram-positive pathogen causing septicaemia and meningitis in pigs and humans. However, how S. suis maintains a high bacterial load in the blood and brain is poorly understood. In this study, we found that a small RNA rss03 is predominantly present in S. suis, Streptococcus parasuis, and Streptococcus ruminantium, implying a conserved biological function. rss03 with a size of 303 nt mainly exists in S. suis sequence type (ST) 1 and epidemic ST7 strains that are responsible for human infections in China. Using MS2-affinity purification coupled with RNA sequencing (MAPS), proteomics analysis, and CopraRNA prediction, 14 direct targets of rss03 from an ST7 strain were identified. These direct targets mainly involve substance transport, transcriptional regulation, rRNA modification, and stress response. A more detailed analysis reveals that rss03 interacts with the coding region of glpF mRNA, and unexpectedly rss03 protects glpF mRNA from degradation by RNase J1. The GlpF protein is an aquaporin, contributes to S. suis oxidative stress resistance by H2O2 efflux, and facilitates bacterial survival in murine macrophages RAW264.7. Finally, we showed that rss03 and GlpF are required to maintain a high bacterial load in mouse blood and brain. Our study presents the first sRNA targetome in streptococci, enriches the knowledge of sRNA regulation in streptococci, and identifies pathways contributing to S. suis pathogenesis.
研究领域:
微生物学
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分子生物学
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感染病学
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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
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分子诊断学
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纳米生物学
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2023
·
IF 7.4
Jiajia Li,Shuang Yue,Ziyuan Gao,Wenhua Hu,Zhaoliang Liu,Guoqiang Xu,Zhen Wu,Xumin Zhang,Guolin Zhang,Fuliang Qian,Junhong Jiang,Shuang Yang
Ribonuclease (RNA) modifications can alter cellular function and lead to differential immune responses by acting as discriminators between RNAs from different phyla. RNA glycosylation has recently been observed at the cell surface, and its dysregulation in disease may change RNA functions. However, determining which RNA substrates can be glycosylated remains to be explored. Here, we develop a solid-phase chemoenzymatic method (SPCgRNA) for targeting glycosylated RNAs, by which glycosylated RNA substrates can be specifically recognized. We found the differential N-glycosylation of small RNAs in hTERT-HPNE and MIA PaCa-2 cancer cells using SPCgRNA. RNA-Seq showed that the changes in glyco-miRNAs prepared from SPCgRNA were consistent with those of traditional methods. The KEGG signaling pathway analysis revealed that differential miRNA glycosylation can affect tumor cell proliferation and survival. Further studies found that NGI-1 significantly inhibited the proliferation, migration, and circulation of MIA PaCa-2 and promoted cell apoptosis. In addition, β-1,4-galactosyltransferase 1 (B4GALT1) not only affected the expression level of glycosylated miRNAs hsa-miR-21-5p but also promoted cell apoptosis and inhibited the cell cycle possibly through the p53 signaling pathway, while B4GALT1 and p53 were also affected following the hsa-miR-21-5p increase. These results suggest that B4GALT1 may catalyze miRNAs glycosylation, which further promotes cancer cell progression.
研究领域:
分子生物学
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生物化学
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肿瘤学
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产品关联
DOI
PubMed
高影响因子
ACS Sensors
·
2024
·
IF 8.2
Rongrong Huang,Peipei Wan,Shengjie Hu,Chenyang Zhang,Wenjun Miao
MicroRNAs (miRNAs) in dermal interstitial fluid (ISF) have recently been recognized as clinically promising biomarkers for the diagnosis and prognosis of cancer. However, the detection poses significant challenges, primarily due to the low abundance of miRNAs and the limitations of current sampling techniques. To address this issue, we develop novel porous microneedles (PMNs) array-based sensor composed of poly(vinyl alcohol) porous hydrogel and DNA-templated silver nanoclusters (AgNCs) to facilitate the enrichment and highly sensitive detection of ISF miRNA. Leveraging the capillary action facilitated by its unique porous structure and the swelling properties of the hydrogel, the PMNs array can efficiently extract 2.7 ± 0.3 mg of ISF within 5 min. Additionally, the interconnected pores within the PMNs array contribute to an increased specific surface area, thereby offering a convenient platform for the decoration of DNA-templated AgNCs. The immobilized large amount of AgNCs effectively capture the target miRNA from the extracted ISF, resulting in miRNA-induced fluorescence quenching of AgNCs. Subsequently, the introduction of the duplex-specific nuclease leads to the cleavage of DNA in DNA-RNA heteroduplexes, which release miRNA to interact with other AgNCs. This process of target recycling triggers a further reduction in fluorescence intensity, thereby enabling sensitive detection of the low-abundant miRNA down to 1.6 pM. Both in vitro and in vivo experiments validate the efficacy of the AgNCs immobilized PMNs array for the detection of miRNA biomarkers in ISF within minutes. These results indicate that the proposed PMNs array-based sensor holds great potential for the development of noninvasive personalized diagnostic strategies.
研究领域:
生物医学工程
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分子诊断
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纳米技术
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Chunrong Li,Yanfei Zhang,Jun Chen,Liping Liu,Huihui Yang,Si-Yang Liu,Yuzhi Xu,Zong Dai,Xiaoyong Zou
Nanomaterial-based on-off-on fluorescence sensing strategies are significant particularly in intracellular nucleic acids imaging assay. There still remains challenge to rationally balance fluorescence quenching efficiency and recovery dynamics. We assume that the performance of on-off-on fluorescence sensing strategy can be fundamentally improved on small zero-dimensional (0D) nanomaterial with precisely modulated surface charge. For a proof-of-concept demonstration, silicon nanoparticle (SiNP) with ~4 nm was synthesized and used as the quencher model, of which the surface charge density was modulated by modification of triphenylphosphonium (TPP). The influence of particle size, surface charge and charge density of the nanomaterials on sensing performance was systematically investigated. The strategy showed a low limit of detection (LOD) as 26 pM for target model miR-494, which is one of the lowest in nanomaterial-based on-off-on sensing platforms. And the LOD is even comparable to amplification-based methods in a greatly shortened assay time (2.5 h). The miR-494 expresses in cancerous and normal living cells of human cervical carcinoma (HeLa), human lung carcinoma (A549), human breast cancer (MCF-7), and normal human mammary epithelial (MCF-10A) cells were imaged and localized with significantly improved sensitivity and specificity. These excellent performances insure it a promising candidate as convenient and non-enzymatic sensing platform for miRNA-associated disease detection and early diagnosis.
研究领域:
生物化学
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分子生物学
·
医学诊断
✓ 已验证
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2015
·
IF 6.409
Chan Song,Guan-Yao Wang,De-Ming Kong
This work investigated the interactions of α-Fe 2 O 3 nanoparticles (NPs) with different structural nucleic acids and their fluorescence quenching ability towards fluorophore-labelled nucleic acid probes . Different from bulk α-Fe 2 O 3 samples, nanoscale α-Fe 2 O 3 particles exhibit the unique properties of strong adsorption and fluorescence quenching to fluorophore-labelled single-stranded DNA (ssDNA) probes. Based on these findings, a facile fluorescence method was developed for versatile quantification of nucleic acids. The size scale of NPs makes a significant impact on this sensing platform. Better selectivity was given by bigger NP (50–100 nm)-based nucleic acid-sensing platform compared with smaller NP (30 nm)-based one. In the 50–100 nm α-Fe 2 O 3 NP-based sensing platform, single nucleotide mismatch or single base-pair mismatch can even be effectively discriminated. The targets of micro-RNA (miRNA), ssDNA and double-stranded DNA (dsDNA) are sensitively detected with detection limits of 0.8 nM, 1.1 nM and 0.64 nM ( S / N =3), respectively. Significantly, α-Fe 2 O 3 NPs possess different affinities towards ssDNA probes with different lengths, and can be used as a universal quencher for ssDNA probes labelled with different fluorescent dyes . On the basis of these properties, the pristine α-Fe 2 O 3 NPs hold the potential to be widely utilized in the development of novel biosensors with signal amplification or simultaneous multiple target detection strategies.
研究领域:
生物化学
·
分子生物学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
CHEMBIOCHEM
·
2020
·Vol. 21
·期 18
·2667-2675
·
IF 2.576
Mengjia Xu,Pan Fu,Shu Xing,Yang Zhao,Chao Zhao
Graphical Point-of-care cancer catching ? A novel PNA-DNA 2 triplex consisting of a homopurine PNA strand and two homopyrimidine DNA strands is reported for the first time. On the basis of this phenomenon, a simple, inexpensive colorimetric PNA-DNA 2 triple-helix molecular switch-based sensing platform for sensitive and selective detection of miRNAs from cancer cells is demonstrated. Peptide nucleic acids (PNAs), the synthetic DNA mimics that can bind to oligonucleotides to form duplexes, triplexes, and quadruplexes, could be advantageous as probes for nucleic acid sequences owing to their unique physicochemical and biochemical properties. We have found that a homopurine PNA strand could bind to two homopyrimidine DNA strands to form a PNA-DNA 2 triplex. Moreover, the cyanine dye DiSC 2 (5) could bind with high affinity to this triplex and cause a noticeable color change. On the basis of this phenomenon, we have designed a label-free colorimetric sensing platform for miRNAs from cancer cells by using a PNA-DNA 2 triple-helix molecular switch (THMS) and DiSC 2 (5). This sensing platform can detect miRNA-21 specifically with a detection limit of 0.18 nM, which is comparable to that of the THMS-mediated fluorescence sensing platform. Moreover, this colorimetric platform does not involve any chemical modification or enzymatic signal amplification, which boosts its applicability and availability at the point of care in resource-limited settings. The universality of this approach can be simply achieved by altering the sequences of the probe DNA for specific targets.
研究领域:
分子生物学
·
医学诊断
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分析化学
✓ 已验证
DOI
高影响因子
ANALYTICAL CHEMISTRY
·
2025
·
IF 6.7
Yanming Lai,Wei Xiao,Zhiwei Lei,Huiping Long,Jiting Deng,Yiyu Wang,Lu Tao,Xingxing Liu,Jinjun Wu,Qiwei Zhang,Donglin Cao,Heng Xiao
Exosomal microRNAs (miRNAs) serve as dependable and noninvasive biomarkers for early cancer diagnosis. However, the accurate and feasible detection of exosomal miRNAs is often hindered by their low abundance and the requirement of specialized equipment for miRNA detection. In this study, we present a novel approach, termed primer exchange reaction-based fluorescence emission with cascade RNA aptamers transcription (PERFECT) for the highly sensitive detection of exosomal miRNA. The design of this method involves the selective interaction of a DNA probe with the target miRNA, leading to its activation. Once activated, isothermal signal amplification and RNA aptamer transcription are initiated, resulting in an amplified fluorescent signal within 90 min. This method achieves a detection limit of 2.2 fM at 37 °C and 2.7 fM at room temperature (25 °C). We used the PERFECT technology to analyze miR-21 expression levels in cell extracts, cell-derived exosomes, and human plasma-derived exosomes, achieving a diagnostic accuracy of 93.6% in distinguishing hepatocellular carcinoma (HCC) patients. Overall, this study highlights its broad range of detection temperature, simplicity of the detection process, and strong potential for clinical application, rendering it a promising tool for early cancer diagnosis.
研究领域:
分子生物学
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医学诊断
·
癌症研究
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产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
·
2018
·
IF 8.097
Qian Zhao,Jiafang Piao,Weipan Peng,Yang Wang,Bo Zhang,Xiaoqun Gong,Jin Chang
Identifying the microRNA (miRNA) expression level can provide critical information for early diagnosis of cancers or monitoring the cancer therapeutic efficacy. This paper focused on a kind of gold-nanoparticle-coated polystyrene microbeads (PS@Au microspheres)-based DNA probe as miRNA capture and duplex-specific nuclease (DSN) signal amplification platform based on an RGB value readout for detection of miRNAs. In virtue of the outstanding selectivity and simple experimental operation, 5′-fluorochrome-labeled molecular beacons (MBs) were immobilized on PS@Au microspheres via their 3′-thiol, in the wake of the fluorescence quenching by nanoparticle surface energy transfer (NSET). Target miRNAs were captured by the PS@Au microspheres-based DNA probe through DNA/RNA hybridization. DSN enzyme subsequently selectively cleaved the DNA to recycle the target miRNA and release of fluorophores, thereby triggering the signal amplification with more free fluorophores. The RGB value measurement enabled a detection limit of 50 fM, almost 4 orders of magnitude lower than PS@Au microspheres-based DNA probe detection without DSN. Meanwhile, by different encoding of dyes, miRNA-21 and miRNA-10b were simultaneously detected in the same sample. Considering the ability for quantitation, high sensitivity, and convenient merits, the PS@Au microspheres-based DNA probe and DSN signal amplification platform supplied valuable information for early diagnosis of cancers.
研究领域:
生物化学
·
分子生物学
·
医学诊断
✓ 已验证
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）