ANALYTICAL CHEMISTRY
·
2026
·Vol. 98
·期 31
·23119-23129
·
IF 7.3
Zhang Lei,Liao Yuqin,Zhu Lvyun,Li Yingying,Yu Lilei,Yang Yanbing,Yuan Quan
Near-infrared II (NIR-II, 1000–1700 nm) fluorescence
imaging
enables deep-tissue analysis, but conventional “always-on”
probes suffer from high background that compromises the signal-to-background
ratio for low-abundance targets. Activatable probe strategies could
mitigate this issue, yet most existing designs rely on fluorescence
resonance energy transfer with acceptors confined to the visible region,
hampering efficient signal quenching in the NIR-II window. Here, we
present a generalizable strategy based on morphology-engineered nanometal
surface energy transfer (NSET) to construct a NIR-II core–satellite
nanoprobe for intracellular miR-21 detection. By systematically tuning
the branch aspect ratio of gold nanostars (AuNSs), their localized
surface plasmon resonance (LSPR) absorption can be red-shifted to
achieve precise spectral overlap with the NIR-II emission of Ag 2 S quantum dots. Guided by DNA self-assembly, an AuNS@Ag 2 S core–satellite structure with controllable interparticle
spacing is established, enabling an initial fluorescence quenching
efficiency of up to 88%. Upon target recognition, miR-21-triggered
strand displacement reaction induces the dissociation of Ag 2 S quantum dots from the AuNS surface, resulting in a pronounced NIR-II
fluorescence “turn-on” response. The nanoprobe exhibits
exceptional performance, including a low detection limit of 75.8 pM,
robust linearity, single-base mismatch discrimination capability,
and enables visualization of differential miR-21 expression in living
cells. Notably, high-contrast NIR-II signals are retained at penetration
depths of up to 15 mm in tissue-mimicking experiments. This work establishes
an NSET-based analytical platform through precise spectral matching
and rational structural engineering, providing a robust strategy for
high signal-to-background biosensing and in situ analysis of low-abundance
miR-21 in deep tissues.
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MICROCHEMICAL JOURNAL
·
2025
·
IF 5.1
Tingyuan Pang,Shuting Ouyang,Jia Li,Xiaorui Liu,Wenying Shu,Wenhui Gao,Junren Li,Qi Song,Jianing Huang,Jun Yang,Xiaoshun Jian,Xiaole Wang,Meimei Zhang
Cervical cancer is one common cause of cancer relative death in women worldwide, while microRNA-21 (miR-21) can serve as one upregulated miRNA in cervical cancer for early diagnosis. Chemiluminescence is a powerful assaying method, but balancing the sensitivity and selectivity is often difficult at present. For this problem, a photocatalytic chemiluminescent biosensor was developed to homogeneously assay miR-21 with good performances. Based on a base stacking principle, cooperative hybridization is employed in double DNA probes for biosensor design. After DNA probe hybridization, photocatalytic DNA-dye complex can induce luminol’s chemiluminescence signals via several processes. Here the double-probe design results in the high sensitivity to assay miR-21 with a detection limit in pM level. Meanwhile, the short probe design and cooperative principle guarantee a good selectivity to discriminate other nucleotide variants. For comparison, both a single short and long probes were also studied to show the advantages of our biosensor in assay performances. So this photocatalytic chemiluminescent biosensor via cooperative hybridization is an ideal method to assay miR-21, showing a good potential for cancer early diagnosis.
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Theranostics
·
2018
·
IF 8.537
Shulian Bai,Bangtian Xu,Yongcan Guo,Juhui Qiu,Wen Yu,Guoming Xie
While detection of microRNA with or without signal amplification is highly informative, nanosensors with high specificity for cell-specific RNA detection are rare. Methods: In this study, a tetrahedral DNA nanostructure (TDN) with a specific function was combined with gold nanoparticles (Au-NP) possessing fluorescence quenching effects and a large surface area to fabricate a fluorescence resonance energy transfer based nanosensor (Au-TDNN). The presence of miR-21 (target) can separate the fluorescent dye-labeled detection probe on Au-TDNNs from Au-NPs, which separates the donor and acceptor, thus inducing an intensive fluorescence signal. High specificity for discerning point mutation targets was achieved by rationally designing the nucleic acid strand displacement reaction to occur spontaneously with ΔG 0 ≈ 0 based on thermodynamic parameters; under this condition, slight thermodynamic changes caused by base mismatch exert significant effects on hybridization yield. Results: Chemically synthesized DNA of three single-base-changed analogues of target, let-7d, and miR-200b were tested. A discrimination factor (DF) of 15.4 was produced by the expected detection probe on Au-NPs for proximal single-base mismatch. As the control group, the DF produced by an ordinary detection probe on Au-NPs only reached 2.4. The feasibility of the proposed strategy was also confirmed using hepatocyte cancer cells (HepG2). Conclusion: This improved nanosensor opens a new avenue for the specific and easy detection of microRNA in live cells.
研究领域:
分子生物学
·
纳米医学
·
生物传感技术
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SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2026
·
IF 4.8
Tingting Guo,Chang Zuo,Ping Bai,Lingjing Hu
A novel multi-functional DNA tetrahedron-based target-induced cascaded cyclic catalytic amplification (TICA) assay was developed for the sensitive detection of microRNA-21 (miR-21) in prostate cancer. This strategy ingeniously integrates two distinct DNA tetrahedron with specific functions (catalytic tetrahedron (Cat-DT) and signal DNA tetrahedron (Sig-DT)) to create a self-sustaining amplification circuit. The process is initiated when miR-21 triggers the release of a DNAzyme enzymatic strand (E-DNA), which converts Cat-DT into tri-branched DNA nanostructures (TB-DNA). Critically, each TB-DNA acts as a cascaded amplifier, bearing three E-DNA units that drive the exponential generation of more catalytic sites. Furthermore, the system employs an ingenious feedback loop: the TB-DNA/Sig-DT interaction produces a fluorescent signal while regenerating a miR-21-mimetic strand to perpetuate the catalytic cycle. This “target → catalyst → new target” paradigm enables ultra-efficient amplification, achieving a wide linear range from 600 fM to 100 nM and a low detection limit of 400 fM for miR-21. Notably, the TICA strategy significantly shortens the total reaction time to 80 min, demonstrating great potential for rapid and sensitive clinical diagnostics.
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Scientific Reports
·
2017
·Vol. 7
·期 1
·1-12
·
IF 4.259
Liu Liang,Xu Qingqing,Hao Shuai,Chen Yun
MicroRNAs (miRNAs) play a vital role in regulating gene expression and are associated with a variety of cancers, including breast cancer. Their distorted and unique expression is a potential marker in clinical diagnoses and prognoses. Thus, accurate determination of miRNA expression levels is a prerequisite for their applications. However, the assays currently available for miRNA detection typically require pre-enrichment, amplification and labeling steps, and most of the assays are only semi-quantitative. Therefore, we developed a quasi-direct liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based targeted proteomics approach to quantify target miRNA by innovatively converting the miRNA signal into the mass response of a reporter peptide via a covalently immobilized DNA-peptide probe. Specifically, the probe containing the targeted proteomics-selected substrate/reporter peptide, GDR AVQLGVDPFR/AVQLGVDPFR, and the DNA sequence complementary to the target miRNA (i.e., miR-21) was first immobilized on APMTS modified silica nanoparticles using PDITC. After the immobilized probe was recognized and hybridized with the target miRNA, the excess probe was degraded using MBN and followed by a trypsin digestion of the hybrids. The reporter peptide was released and quantified using LC-MS/MS. The obtained LOQ was 5 pM. Finally, the developed assay was used for the quantitative analysis of miR-21 in breast cells and tissue samples.
研究领域:
分子生物学
·
癌症研究
·
蛋白质组学
·
分析化学
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ACTA BIOCHIMICA POLONICA
·
2021
·Vol. 68
·期 4
·725-732
·
IF 2.149
Ziyan Wu,Zhong Zhou,Weiguo Zhang,Yi Yu
MicroRNA (miR)-21 has been found to be overexpressed in osteosarcoma (OS). The aim of the present study was to investigate the effect of miR-21-5p on the Warburg effect and stemness maintenance in OS cells and its potential molecular mechanism. Herein, miR-21-5p was overexpressed or inhibited in MG-63 cells via transfection with mimics or inhibitors. The effect of miR-21-5p on cell viability, apoptosis, Warburg effect and stemness maintenance were explored in OS cells. The results demonstrated that miR-21-5p inhibition suppressed MG-63 cell viability and enhanced their apoptosis. Additionally, miR-21-5p inhibition attenuated the stemness maintenance of MG-63 cells, as demonstrated by the reduced proportion of CD133-positive MG-63 cells, the decrease in tumorsphere formation capacity, and the downregulation of Sox2, Oct4, and Nanog proteins. Moreover, miR-21-5p inhibition suppressed the Warburg effect in MG-63 cells, as indicated by the decrease in glucose uptake, lactic acid production, and ATP level and the downregulation of proteins involved in the Warburg effect (GLUT1, LDHA, HK2, and PKM2). Furthermore, the results suggested that the effect of miR-21-5p suppression on stemness and the Warburg effect may be associated with the decreased activity of the Wnt/β-catenin pathway in OS cells. Our findings suggest a novel potential biomarker for OS therapy.
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ANALYTICAL CHEMISTRY
·
2015
·
IF 5.636
Feifei Xu,Ting Yang,Yun Chen
The distorted and unique expression of microRNAs (miRNAs) in cancer makes them an attractive source of biomarkers. However, one of prerequisites for the application of miRNAs in clinical practice is to accurately profile their expression. Currently available assays normally require pre-enrichment, amplification, and labeling steps, and most of them are semiquantitative. In this study, we converted the signal of target miR-21 into reporter peptide by a DNA-peptide probe and the reporter peptide was ultimately quantified using LC-MS/MS-based targeted proteomics. Specifically, substrate peptide GDKAVLGVDPFR containing reporter peptide AVLGVDPFR and tryptic cleavage site (lysine at position 3) was first designed, followed by the conjugation with DNA sequence that was complementary to miR-21. The newly formed DNA-peptide probe was then hybridized with miR-21, which was biotinylated and attached to streptavidin agarose in advance. After trypsin digestion, the reporter peptide was released and monitored by a targeted proteomics assay. The obtained limit of quantification (LOQ) was 1 pM, and the detection dynamic range spanned ∼5 orders of magnitude. Using this assay, the developed quasi-targeted proteomics approach was applied to determine miR-21 level in breast cells and tissue samples. Finally, qRT-PCR was also performed for a comparison. This report grafted the strategy of targeted proteomics into miRNA quantification.
研究领域:
癌症生物学
·
分子生物学
·
蛋白质组学
·
分析化学
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Small
·
2018
·Vol. 14
·期 10
·1703858
·
IF 9.598
Keying Zhang,Lin Yang,Feng Lu,Xingcai Wu,Jun-Jie Zhu
Here, a sensitive and universal noncoding RNA (ncRNA) upconversion sensing nanoplatform is developed. Gold nanoparticles bearing one hairpin DNA (Hp) molecule are conjugated to the linker DNA modified NaYF 4 :Yb, Er@NaYF 4 upconversion nanoparticles by DNA hybridization, leading to quenching of the upconversion emission through fluorescence resonance energy transfer. A signal DNA (SDNA) sequence is designed to open Hp, recovering the upconversion emission. To achieve universality and high sensitivity of the nanoprobe, an exonuclease III (Exo III)-assisted cycling amplification strategy is introduced. A multifunctional hairpin DNA (mHp) containing ncRNA recognition sequence and SDNA sequence is designed to recognize ncRNA and trigger Exo III as a biocatalyst to stepwise disintegrate itself, releasing both ncRNA and SDNA. The released ncRNA can be reused to release more SDNA, which greatly improves the sensing sensitivity. By changing the recognition portion of mHp, various ncRNA can be detected. The sensitive detection of both homeobox (HOX) transcript antisense RNA segment and miR-21 is achieved with this novel strategy, even in human serum, indicating the universality and sensitivity of the proposed strategy. Additionally, the expression level of miR-21 in human breast cancer cell (MCF-7) lysate is successfully measured, suggesting its potential in clinical diagnosis.
研究领域:
生物技术
·
纳米技术
·
癌症诊断
·
分子生物学
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Polymer Chemistry
·
2016
·Vol. 7
·期 26
·4352-4366
·
IF 5.687
Changyu He,Zhen Zhang,Qinglai Yang,Qing Chang,Zhifeng Shao,Bing Gong,Yu-Mei Shen,Bingya Liu,Zhenggang Zhu
A reductive tri-block copolymer PEG-SS-PLA-SS-PEI with a double disulphide linkage in the backbone directed by H-bonding association was synthesized and self-assembled into cationic polymeric nanomicellar particles for in vivo antimiRNA delivery with an average diameter of 68 nm and a zeta potential of approximately 39 mV. The chemical structure of the copolymer was well characterized by 1H NMR and GPC. The cationic polymeric nanomicellar particles could be unpacked in an intracellular reductive environment (GSH) leading to the release of encapsulated antimiRNA. MTT assays in vitro showed no significant cytotoxicity of SGC7901 gastric cancer cells incubated with PEG-SS-PLA-SS-PEI micelles. The in vitro study indicated that the micelle-based antimiR-21 delivery system could effectively facilitate cellular uptake and greatly down-regulate the expression level of miR-21 in SGC7901 cell lines, which was comparable to Lipofectamine™ 2000. The down regulation of miR-21 remarkably induced apoptosis, suppressed the tumor cell migration and invasion, and increased the expression of target genes such as phosphatase and tensin homolog deleted on chromosome ten (PTEN) and Programmed Cell Death Protein 4 (PDCD4). More importantly, the in vivo systemic administration of the micelles/antimiR-21 complex in a gastric cancer model significantly inhibited tumor growth and increased the expression of target genes. The nanoparticle based on the PEG-SS-PLA-SS-PEI copolymer would be a safe and efficient carrier for delivery of therapeutic antimiRNA, which shows a prospective therapy method in gastric cancer.
研究领域:
纳米技术
·
药学
·
肿瘤学
·
分子生物学
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BIOSENSORS & BIOELECTRONICS
·
2023
·
IF 12.6
Jiwei Wang,Jingjing Fu,Han Chen,Ali Wang,Yuting Ma,Hanrong Yan,Yuting Li,Dehong Yu,Fenglei Gao,Shibao Li
Accurate quantitative, in situ and temporal tracking imaging of tumor-associated miRNAs in living cells could provide a basis for cancer diagnosis and prognosis. In this strategy, a surface-enhanced Raman scattering (SERS)-fluorescence (FL) dual-spectral sensor (DSS) was constructed based on the nanoscale photophysical properties of AuNPs , mediated by functionalized DNA , to achieve rapid imaging of FL and accurate SERS quantification of intracellular miRNAs. The dual-spectrum sensor in the strategy is highly sensitive, specific and reproducibly stable. The LOD values of the dual spectra were 3.58 pM (SERS) as well as 11.8 pM (FL) with RSD values less than 2.69%. The bispectral sensor self-assembled into a trimer by the lapidation of Y-type DNA under the excitation of the target, generating a stable enhanced electric field coupling; and selected adenine located in the enhanced electric field as the reporter molecule, simplifying the labeling process and variables of the Raman reporter molecule, distinguishing it from other traditional methods. This strategy successfully achieved accurate tracking and quantification of miR-21 in cancer cells and showed good stability in the cells. The reported probes are potential tools for reliable monitoring of biomolecular dynamics in living cells.
研究领域:
纳米技术
·
生物医学工程
·
分子生物学
·
癌症研究
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ANALYST
·
2024
·
IF 3.6
Haiying Cai,Peiting Dong,Xiuping Li,Lulu Wang,Tao Li
It is known that the abnormal expression of specific cellular miRNAs is closely related to cell apoptosis, and so monitoring the level change of these miRNAs can in principle be used to evaluate the process of apoptosis stimulated by drugs. Towards this goal, here we construct an ultrasensitive electrochemiluminescence (ECL) nanoplatform via the target miRNA-triggered immobilization of spherical nucleic acid enzymes (SNAzymes) onto tetrahedral DNA nanostructures on the electrode surface, which catalyzes the luminol–H2O2 reaction to output an ECL signal. This enables the sensitive and specific detection of two apoptosis-related miRNAs, miR-21 and miR-133a, with a detection limit of 33 aM. Furthermore, we employed the developed ECL nanoplatform to monitor the levels of these two miRNAs inside cancer cells stimulated by DOX, showing that the level of miR-21 decreases, while that of miR-133a increases in the early apoptotic cells. This difference highlights the distinct roles of the two target miRNAs, where miR-21 promotes the early apoptosis of cancer cells, whereas miR-133a suppresses it, providing new insight into cell physiological processes.
研究领域:
生物技术
·
纳米技术
·
分子生物学
·
癌症研究
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ANALYTICAL CHEMISTRY
·
2022
·
IF 8.008
Yanfei Zhang,Jun Chen,Huihui Yang,Wen Yin,Chunrong Li,Yuzhi Xu,Si-Yang Liu,Zong Dai,Xiaoyong Zou
Sensitive imaging of intracellular microRNA (miRNA) in living cells is of great significance. Isothermal hybridization chain reaction (HCR)-based methods, although have been widely used to monitor intracellular low-abundance miRNA, are still subjected to the challenges of limited signal amplification efficiency and compromised imaging resolution. In this work, we design a light-controlled recruitable HCR (LCR-HCR) strategy that enables us to well overcome these limitations. Exosomes as delivery and recruitment vehicles are modified with three cholesterol-modified hairpins (H1, H2, and H3), in which H1 is for anchoring target miRNA and H2 and H3 with photocleavable linkers (PC-linkers) are designed for spatiotemporal HCR. By controllably releasing probes with high local concentrations to efficiently trigger HCR and further recruiting the generated double-stranded DNA (dsDNA) polymers instead of dispersion in the cytoplasm, the LCR-HCR method can significantly improve the imaging contrast by confining all of the reactants on exosome vehicles. For a proof-of-concept demonstration, the miR-21 was analyzed by LCR-HCR with a limit of detection (LOD) down to 3.3 pM (corresponding to 165 amol per 50 μL) in vitro and four times higher response than traditional HCR in vivo. In general, the LCR-HCR method provides a powerful tool for sensitive miRNA imaging in living cells and cancer diagnosis.
研究领域:
生物技术
·
分子生物学
·
癌症研究
·
细胞生物学
·
纳米技术
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