Frontiers in Bioengineering and Biotechnology
·
2023
·
IF 5.7
Yu Xiongtao,Bai Silan,Wang Lishi
In this work, gold nanoparticles@Ti3C2 MXenes nanocomposites with excellent properties were combined with toehold-mediated DNA strand displacement reaction to construct an electrochemical circulating tumor DNA biosensor. The gold nanoparticles were synthesized in situ on the surface of Ti3C2 MXenes as a reducing and stabilizing agent. The good electrical conductivity of the gold nanoparticles@Ti3C2 MXenes composite and the nucleic acid amplification strategy of enzyme-free toehold-mediated DNA strand displacement reaction can be used to efficiently and specifically detect the non-small cell cancer biomarker circulating tumor DNA KRAS gene. The biosensor has a linear detection range of 10 fM −10 nM and a detection limit of 0.38 fM, and also efficiently distinguishes single base mismatched DNA sequences. The biosensor has been successfully used for the sensitive detection of KRAS gene G12D, which has excellent potential for clinical analysis and provides a new idea for the preparation of novel MXenes-based two-dimensional composites and their application in electrochemical DNA biosensors.
研究领域:
生物医学工程
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分子诊断学
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
Pharmaceutics
·
2021
·Vol. 13
·期 9
·1428
·
IF 6.321
Dandan Luo,Xiaochun Xu,M. Zubair Iqbal,Qingwei Zhao,Ruibo Zhao,Jabeen Farheen,Quan Zhang,Peiliang Zhang,Xiangdong Kong
Pancreatic carcinoma (PC) is greatly induced by theKRASgene mutation, but effective targeted delivery for gene therapy has not existed. Small interfering ribonucleic acid (siRNA) serves as an advanced therapeutic modality and holds great promise for cancer treatment. However, the development of a non-toxic and high-efficiency carrier system to accurately deliver siRNA into cells for siRNA-targeted gene silencing is still a prodigious challenge. Herein, polyethylenimine (PEI)-modified hydroxyapatite (HAp) nanoparticles (HAp-PEI) were fabricated. The siRNA of theKRASgene (siKras) was loaded onto the surface of HAp-PEI via electrostatic interaction between siRNA and PEI to design the functionalized HAp-PEI nanoparticle (HAp-PEI/siKras). The HAp-PEI/siKras was internalized into the human PC cells PANC-1 to achieve the maximum transfection efficiency for active tumor targeting. HAp-PEI/siKras effectively knocked down the expression of theKRASgene and downregulated the expression of the Kras protein in vitro. Furthermore, the treatment with HAp-PEI/siKras resulted in greater anti-PC cells’ (PANC-1, BXPC-3, and CFPAC-1) efficacy in vitro. Additionally, the HAp-PEI exhibited no obvious in vitro cytotoxicity in normal pancreatic HPDE6-C7 cells. These findings provided a promising alternative for the therapeutic route of siRNA-targeted gene engineering for anti-pancreatic cancer therapy.Keywords:hydroxyapatite;siRNA delivery;pancreatic cancer cells;KRASgene;gene silence;anticancer
研究领域:
纳米技术
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癌症生物学
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基因治疗
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药学
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产品关联
DOI
PubMed
高影响因子
MICROCHIMICA ACTA
·
2024
·Vol. 191
·期 6
·1-10
·
IF 5.3
Wang Lanyue,Bu Sisi,Xu Shijie,Huang Tuo,Yang Fang,Tan Qianglong,Deng Minxin,Xie Wenlin,Cai Bobo,Chen Jian
Single nucleotide polymorphism (SNP) biosensors are emerging rapidly for their promising applications in human disease prevention diagnosis, treatment, and prognosis. However, it remains a bottleneck in equipping simple and stable biosensors with the traits of high sensitivity, non-enzyme, and low cost. Double base mismatches mediated chain displacement reactions have attracted fascinating advantages of tailorable thermodynamics stability, non-enzyme, and excellent assembly compliance to involvement in SNP identification. As the base mismatch position and amount in DNA sequence can be artificially adjusted, it provides plenty of selectivity and specificity for exploring perfect biosensors. Herein, a biosensor with double base mismatches mediated catalytic hairpin assembly (CHA) is designed via one base mismatch in the toehold domain and the other base mismatch in the stem sequence of hairpin 1 (H1) by triggering CHA reaction to achieve selective amplification of the mutation target (MT) and fluorescence resonance energy transfer (FRET) effect that is composed of Cy3 and Cy5 terminally attached H1 and hairpin 2 (H2). Depending on the rationally designed base mismatch position and toehold length, the fabricated biosensors show superior SNP detection performance, exhibiting a good linearity with high sensitivity of 6.6 fM detection limit and a broad detection abundance of 1%. The proposed biosensor can be used to detect the KRAS mutation gene in real samples and obtain good recoveries between 106 and 116.99%. Remarkably, these extendible designs of base mismatches can be used for more types of SNP detection, providing flexible adjustment based on base mismatch position and toehold length variations, especially for their thermodynamic model for DNA-strand displacement reactions. Graphical Abstract
研究领域:
分子生物学
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生物传感技术
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医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2022
·
IF 9.221
Peilin Wang,Zizhun Wang,Zhenrun Li,Yuan Wang,Qiang Ma
In this work, a novel electrochemiluminescence (ECL) biosensor was fabricated to detect the mutant KRAS gene in the tumor tissues. Firstly, a new kind of chitosan-based dots was prepared and regulated by phenylboronic acid (PBA) as ECL nanoprobe. Furthermore, the electrochemically deposited Ag structures with controllable morphology have been developed on the electrode. The detailed electrochemical deposition condition of Ag structure has been investigated deeply. Due to the synergistic effect of the large active surface area and the high conductivity , the layered Ag flower-like structure with branches can significantly enhance the ECL intensity of CS-PBA dots by 12.1 times. Finally, the toehold mediated strand displacement strategy was employed in the ECL biosensor to quantitatively detect mutant KRAS gene with the range of 0.01 pM to 10 nM. The limit of detection was 3.3 fM. This biosensor has been used successfully to analyze the target DNA in the tumor tissues of actual cancer patients . The results showed the novel sensing platform possessed great potential for clinical analysis.
研究领域:
生物医学工程
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分析化学
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分子生物学
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癌症研究
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2025
·
IF 7.7
Li Meng,Mingzhong Li,Zhenyu Xu,Tianlan Chen,Meiwan Chen,Joana Vaz Pinto,Bruno Veigas,Man-Kay Law
Precise temperature control and measurement are crucial for in vitro nucleic acid denaturation, especially in melting curve analysis (MCA). However, the inherent cumbersome multi-step calibration process required to obtain an accurate spatial temperature gradient can be a major limitation for the study of MCA on existing microfluidic chips. This work proposes to obtain a highly linear temperature gradient with a functional substrate for achieving high-resolution MCA on a digital microfluidics (DMF) platform. Taking advantage of the high thermal conductivity of aluminum, we employ a thin trapezoidal aluminum plate as the DMF substrate to compensate for the uneven heat transfer caused by temperature differences at different substrate positions and the ambient environment. We further pattern a banded superhydrophilic thread on the superhydrophobic surface to passively achieve a liquid thread by driving the mother droplet through the electrowetting on dielectric (EWOD) effect. With a heater and a cooler positioned at the opposite ends of the substrate, we can create a highly linear (simulated R 2 = 1 and experimental R 2 = 0.9998) temperature gradient from 40°C to 90°C across the liquid thread, requiring only temperature calibration at the two end points. The elimination of laborious temperature calibration steps can effectively enable rapid, continuous and reliable nucleic acid amplification as well as MCA on DMF. The high temperature accuracy also ensures simultaneous multiple hybrid DNA detection for rapid one-time discrimination of single nucleotide mutations in KRAS gene targets. This innovation underscores the DMF platform's potential in the realms of biomedical research and clinical diagnostics.
研究领域:
生物医学工程
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微流控技术
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分子诊断学
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生物化学
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遗传学
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芯片实验室技术
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临床诊断学
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产品关联
DOI
高影响因子
ANALYTICAL SCIENCES
·
2018
·Vol. 35
·期 4
·441-448
·
IF 1.355
Wang Xiaojing,Yang Mei,Liu Qingyan,Yang Siyi,Geng Xintong,Yang Yixia,Fa Huanbao,Wang Yongzhong,Hou Changjun
In this paper, an ultrasensitive electrochemical biosensor based on carboxylated multi-walled carbon nanotube/molybdenum disulfide composites (MWCNTs-COOH/MoS 2 ) for the detection of KRAS gene is described. An easy, low-cost method, named one-step hydrothermal, was used for the synthesize of MWCNTs-COOH/MoS 2 nanocomposites, and scanning electronic microscopy (SEM), high resolution transmission electron microscopy (HRTEM), Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS) and X-ray diffraction (XRD) were used for characterizing the prepared composites. Furthermore, cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were employed for an electrochemical performance study of this biosensor. Under optimal conditions, the detection limit of target DNA achieved down to 3 fM ( S / N = 3) with high sensitivity; the linear range with the logarithm of the concentrations of target DNA varied from 1.0 × 10 −14 to 1.0 × 10 −7 M. Finally, the practicality of our proposed sensor was verified by a determination of the KRAS gene in human serum samples with good accuracy and high precision due to the excellent conductivity and large active surface area of the MWCNTs-COOH/MoS 2 nanocomposites. This proposed biosensor thus provides a practical method for the rapid and sensitive analysis of gene detection.
研究领域:
纳米技术
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生物传感器
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电化学
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基因检测
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材料科学
✓ 已验证
DOI
PubMed
高影响因子
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