JOURNAL OF ELECTROANALYTICAL CHEMISTRY
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2020
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IF 3.807
Ziheng Hu,Biying Zhao,Ping Miao,Xialing Hou,Feifei Xing,Yingying Chen,Lingyan Feng
Herein, we present one three-way junction (TWJ) DNA based electrochemical biosensor for sensitive microRNAs detection with locked nucleic acid (LNA) as capture probe and enzyme-free redox cycling signal amplification strategy. LNA owns higher binding affinity and thermal stability compared to normal DNA sequence . In the presence of target microRNA (miRNA-21), novel TWJ structure can be specially triggered with the assistant of methylene blue (MB)-tagged DNA signal probe, thus producing an enhanced electrochemical current. Furthermore, a chemical reductant, tris (2-carboxyethyl) phosphine (TCEP), is adopted to lead a relayed electron transport between solution TCEP and TWJ-linked MB tag and amplified whole signal intensity. Based on the selectivity from target-induced LNA/DNA TWJ structure and enzyme-free MB cycling amplification strategy, the designed electrochemical biosensor exhibits sensitive miRNA-21 detection, with a linear range from 100 aM to 100 pM and a low detection limit of 77 aM. In addition, it can successfully discriminate the differences between the miRNA family members, presenting a promising response to the direct determination of target miRNA-21 in real biological fluids. Our work provides an alternative LNA-based electrochemical biosensor way in the area of miRNA diagnostics and clinical analysis without complicated enzyme/nanomaterials-assisted signal amplification.
研究领域:
生物化学
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分子诊断
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分析化学
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产品关联
DOI
高影响因子
ANALYTICAL CHEMISTRY
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2014
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IF 5.825
Zhong Feng Gao,Yu Ling,Lu Lu,Ning Yu Chen,Hong Qun Luo,Nian Bing Li
Although various strategies have been reported for single-nucleotide polymorphisms (SNPs) detection, development of a time-saving, specific, and regenerated electrochemical sensing platform still remains a realistic goal. In this study, an ON–OFF switching of a regenerated biosensor based on a locked nucleic acid (LNA)-integrated and toehold-mediated strand displacement reaction technique is constructed for detection of SNPs. The LNA-integrated and methylene blue-labeled capture probe with an external toehold is designed to switch on the sensing system. The mutant-type DNA probe completes complementary with the capture probe to trigger the strand displacement reaction, which switches off the sensing system. However, when the single-base mismatched wild-type DNA probe is presented, the strand displacement reaction cannot be achieved; therefore, the sensing system still keeps the ON state. This DNA sensor is stable over five reuses. We further testify that the LNA-integrated sequence has better recognition ability for SNPs detection compared to the DNA-integrated sequence. Moreover, this DNA senor exhibits a remarkable discrimination capability of SNPs among abundant wild-type targets and 6000-fold (m/m) excess of genomic DNA. In addition, it is selective enough in complex and contaminant-ridden samples, such as human urine, soil, saliva, and beer. Overall, these results demonstrate that this reliable DNA sensor is easy to be fabricated, simple to operate, and stable enough to be readily regenerated.
研究领域:
分子生物学
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生物传感技术
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遗传学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF CONTROLLED RELEASE
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2025
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IF 11.5
Li Zhou,Fangfang Duan,Yuting Zhang,Yicai Huang,Le Xu,Nan Zhang,Shiming Yang,Yi Ma,Bing Yang,Kai Lu
Hyperpigmentation is a skin disorder characterized by the abnormal accumulation of melanin, in which the melanocortin-1 receptor (MC1R) was recognized as a key target for melanin production. Antisense oligonucleotide (ASO), by targeting mRNA to inhibit protein synthesis, holds a promising gene therapy for hyperpigmentation. However, challenges such as nonspecific activation, off-target toxicity, and limited cellular permeability hinder their broader clinical application. Inspired by the Chinese proverb “Turn an enemy into a friend”, we introduce a dual-gated, ultraviolet (UV)-tyrosinase (TYR) cascade-responsive modified ASO (m-ASO) that enables spatiotemporally controlled silencing of MC1R. The engineered m-ASO is caged via thymidine modification, rendering it inactive until sequentially exposed to exogenous UV and intracellular TYR. UV irradiation, conventionally hyperpigmentation trigger, is repurposed to initiate decaging of m-ASO into an intermediate form, pH-ASO. This intermediate is then enzymatically activated by TYR overexpressed in melanocytes within pigmented tissues, ultimately restoring the native conformation and therapeutic function of the ASO. This “enemy-to-ally” strategy allows for disease-specific, precision activation of ASO therapeutics. Notably, to enhance transdermal delivery, the m-ASO nanocomplex is integrated into a biocompatible, dissolvable microneedle system for painless and localized epidermal administration. The resulting platform demonstrates precise MC1R silencing and effective melanin suppression, while sparing normal melanocytes. Together, this cascade-caged, cell-selective, and minimally invasive strategy presents a novel framework for programmable nucleic acid therapeutics in dermatological and enzyme-overexpressing diseases.
研究领域:
皮肤病学
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基因治疗
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核酸治疗
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药物递送系统
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分子生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
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2021
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IF 6.057
Min-Xi Li,Yao Chen,Zeng-Ping Chen,Ru-Qin Yu
Herein, a label free and sensitive miRNA detection method with enhanced practical applicability was developed based on the locked nucleic acid (LNA) assisted repeated fishing amplification strategy. The working mechanism of the proposed method is as follows: 1) a DNA probe (i.e, L-DNA) with LNA bases is immobilized onto the surface of a gold foil. The L-DNA hybridizes with the 3′ terminus of the first strands of complementary deoxyribonucleic acid (cDNA) of the target miRNA in the test samples; 2) The protruding 5′ terminus of the cDNA serves as a ‘fishhook’ to repeatedly fish the products of a hybridization chain reaction (HCR) out from a ‘reaction tube’; 3) The HCR products can be unloaded from the gold foil into a ‘product tube’ through temperature-controlled dehybridization; 4) The concentration of the target miRNA is determined based on the fluorescence intensity generated by the addition of SYBR-Green I (SG) into the ‘product tube’. The proposed platform was applied to the detection of miRNA-122 in cell lysate samples and obtained quantitative results with accuracy comparable to the quantitative reverse transcription PCR method (qRT-PCR). It is worth pointing out that the proposed platform achieved a limit of detection value of 2.9 fM for miRNA-122 by a simple but effective LNA-assisted repeated fishing amplification strategy instead of complicated enzyme-based amplification techniques. It is reasonable to expect that the proposed method provides a competitive alternative for designing practically applicable, cost-effective and label-free miRNA detection methods.
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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分析化学
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产品关联
DOI
PubMed
高影响因子
ELECTROCHIMICA ACTA
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2012
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IF 3.832
Xiaomeng Meng,Minrong Xu,Jianying Zhu,Huanshun Yin,Shiyun Ai
In this paper, a novel sensitive biosensor was fabricated for DNA detection based on gold nanoparticles (AuNPs) locked nucleic acid modified hairpin DNA probe (LNA-m-HpDNA) and enzymatic signal amplification. LNA-m-HpDNA dually labeled with 3′-thiol and 5′-biotin was efficiently assembled on the electrode via the interaction of thiol and gold. The stem-loop structure of LNA-m-HpDNA was unfolded after hybridization with target DNA, which forced biotin away from the electrode surface. Streptavidin-labeled horseradish peroxidase (Streptavidin-HRP) was subsequently immobilized on the electrode surface via the specific conjugation of biotin and streptavidin. Thereafter, the resulting HRP/target DNA–LNA-m-HpDNA (cDNA–LNA)/AuNPs modified Au electrode was successfully assembled for the construction of the DNA biosensor. Cyclic voltammetry and electrochemical impedance spectroscopy were carried out for the characterization of modified electrodes. Electrochemical reduction signal of benzoquinone was investigated in this work to indirectly analyze DNA hybridization via chronoamperometry. Under optimum conditions, the biosensor showed a good linear relationship between the current value and logarithm of the target DNA concentration ranging from 10 to 1000 pM with a low detection limit of 6.0 pM (S/N = 3). Furthermore, the DNA biosensor exhibited excellent discrimination ability to detect single-base mismatched DNA, three-base mismatched DNA and non-complementary DNA sequence. This strategy could provide a new platform for disease diagnose with high sensitivity.
研究领域:
生物化学
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纳米技术
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分子生物学
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分析化学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）