Analytical Methods
·
2025
·
IF 2.6
Changqing Du,Anran Li,Jinye Qiu,Jiarui Sun,Xiaokun Wang,Wei Xu,Maofeng Zhang,Dapeng Wang
Quantitative mRNA expression levels are essential for differential and clinical diagnoses. In this work, we present an apurinic/apyrimidinic endonuclease 1 (APE1)-assisted SERS sensing platform for rapid and ultrasensitive detection of TK1 mRNA. This platform utilizes a hairpin probe that contains a TK1 mRNA capture sequence along with an apurinic/apyrimidinic (AP) sites, which is specifically recognized and cleaved by APE1. Upon introduction of the target mRNA, hybridization occurs between the mRNA and the AP probe (HAP) within the hairpin structure, forming a doublestranded complex. APE1 then cleaves the HAP at the specific AP sites within the double-stranded complex, leading to a measurable change in the SERS intensity of the FAM-labeled HAP. Based on this principle, the developed SERS sensing platform demonstrates a highly linear response for TK1 mRNA detection across a wider range from 1 fM to 10 nM, with a detection limit (LOD) of 0.2 fM. Moreover, the SERS sensing platform shows great potential for the precise and quantitative detection of TK1 mRNA in human serum, which could be instrumental for mRNA-related research and the early clinical diagnosis of diseases.
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高影响因子
MICROCHIMICA ACTA
·
2023
·Vol. 190
·期 11
·1-9
·
IF 5.7
Wang Jing,Shi Lu,Tang Qiaorong,Zhu Xinyu,Wu Mengmeng,Liu Wei,Li Baoxin,Jin Yan
Nucleic acid lateral flow assay (NALFA) with gold nanoparticles (AuNPs) as colorimetric probes have been extensively adopted for point-of-care testing (POCT). However, the sensitivity of NALFA still needs to be improved. Herein, DNA-gold nanoaggregate (DNA-AuNA) was assembled as a signal amplification probe of NALFA for sensitive detection of tumor marker TK1 mRNA. Four functional oligonucleotides with complementary pairs were assembled to form DNA-AuNA that coupled more AuNPs to improve sensitivity. Thus, the limit of detection (LOD) was 0.36 pM, which is lower than that of conventional AuNPs-based NALFA. Moreover, the bioassay showed good reproducibility, stability, and specificity for detecting TK1 mRNA. The detection of TK1 mRNA in human serum was also satisfactory. Therefore, DNA-AuNA-based NALFA provides a sensitive method for portable detection of TK1 mRNA. Graphical abstract
研究领域:
生物化学
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分子诊断
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纳米医学
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高影响因子
Small
·
2016
·Vol. 12
·期 43
·5944-5953
·
IF 8.315
Qianwen Ding,Qiuqiang Zhan,Xiaoming Zhou,Tao Zhang,Da Xing
Remote optical detection and imaging of specific tumor-related biomarkers and simultaneous activation of therapy according to the expression level of the biomarkers in tumor site with theranostic probes should be an effective modality for treatment of cancers. Herein, an upconversion nanobeacon (UCNPs-MB/Dox) is proposed as a new theranostic nanoprobe to ratiometrically detect and visualize the thymidine kinase 1 (TK1) mRNA that can simultaneously trigger the Dox release to activate the chemotherapy accordingly. UCNPs-MB/Dox is constructed with the conjugation of a TK1 mRNA-specific molecular beacon (MB) bearing a quencher (BHQ-1) and an alkene handle modified upconversion nanoparticle (UCNP) through click reaction and subsequently loading with a chemotherapy drug (Dox). With this nanobeacon, quantitative ratiometric upconversion detection of the target with high sensitivity and selectivity as well as the target triggered Dox release in vitro is demonstrated. The sensitive and selective ratiometric detection and imaging of TK1 mRNA under the irradiation of near infrared light (980 nm) and the mRNA-dependent release of Dox for chemotherapy in the tumor MCF-7 cells and A549 cells are also shown. This work provides a smart and robust platform for gene-related tumor theranostics.
研究领域:
纳米技术
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癌症生物学
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分子影像
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治疗诊断学
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高影响因子
MOLECULES
·
2025
·Vol. 30
·期 17
·3551
·
IF 4.6
Senhao Hu,Wenjing Wang,Yu Zou,Chunmei Li,Hongyan Zou,Chengzhi Huang,Lei Zhan
The development of low-background, facile, and robust fluorescent nanoprobes for imaging and monitoring of intracellular mRNA changes remains a great challenge. Taking advantage of the high fluorescence quenching efficiency of core-shell gold@polydopamine (Au@PDA) nanocomposites and Ca 2+ -promoting DNA adsorption stability, a simple and universal bioconjugate strategy was designed to a construct fluorescent nanoprobe for highly efficient tumor-related mRNA imaging. The fluorescence of Cy5-labeled DNA was quenched up to 92.38% by the AuNP and PDA via nanometal surface energy transfer (NSET) and photoinduced electron transfer (PET), respectively. TK1 mRNA, a biomarker of tumor growth, initiates hybridization and results in fluorescence recovery, which built the foundation for identifying the expression level changes in living cells. More importantly, three kinds of tumor-related mRNA (TK1 mRNA, GalNAc-T mRNA, and C-myc mRNA) can be detected simultaneously with different fluorophore-modified recognition sequences, which can avoid false positive signals and improve the reliability of cancer diagnostic, holding great promise for cancer diagnosis, prognosis, and therapy.
研究领域:
纳米生物技术
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分子成像
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癌症诊断
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生物分析化学
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基因表达监测
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高影响因子