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Photoresponsive CHA-Integrated Self-Propelling 3D DNA Walking Amplifier within the Concentration Localization Effect of DNA Molecular Framework Enables Highly Efficient Fluorescence Bioimaging
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-23 , DOI: 10.1021/acs.analchem.3c04920 Jing-Wei He 1 , Xiaoming Sun 2 , Hong-Wu Tang 3 , Da Liu 1 , Cheng-Yu Li 1
Analytical Chemistry ( IF 6.7 ) Pub Date : 2024-01-23 , DOI: 10.1021/acs.analchem.3c04920 Jing-Wei He 1 , Xiaoming Sun 2 , Hong-Wu Tang 3 , Da Liu 1 , Cheng-Yu Li 1
Affiliation
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While three-dimensional (3D) DNA walking amplifiers hold considerable promise in the construction of advanced DNA-based fluorescent biosensors for bioimaging, they encounter certain difficulties such as inadequate sensitivity, premature activation, the need for exogenous propelling forces, and low reaction rates. In this contribution, a variety of profitable solutions have been explored. First, a catalytic hairpin assembly (CHA)-achieved nonenzymatic isothermal nucleic acid amplification is integrated to enhance sensitivity. Subsequently, one DNA component is simply functionalized with a photocleavage-bond to conduct a photoresponsive manner, whereby the target recognition occurs only when the biosensor is exposed to an external ultraviolet light source, overcoming premature activation during biodelivery. Furthermore, a special self-propelling walking mechanism is implemented by reducing biothiols to MnO2 nanosheets, thereby propelling forces that are self-supplied to a Mn2+-reliant DNAzyme. By carrying the biosensing system with a DNA molecular framework to induce a unique concentration localization effect, the nucleic acid contact reaction rate is notably elevated by 6 times. Following these, an ultrasensitive in vitro detection performance with a limit of detection down to 2.89 fM is verified for a cancer-correlated microRNA biomarker (miRNA-21). Of particular importance, our multiple concepts combined 3D DNA walking amplifier that enables highly efficient fluorescence bioimaging in live cells and even bodies, exhibiting a favorable application prospect in disease analysis.
中文翻译：
光响应 CHA 集成自推进 3D DNA 步行放大器在 DNA 分子框架的浓度定位效应内实现高效荧光生物成像
虽然三维 (3D) DNA 步行放大器在构建用于生物成像的先进的基于 DNA 的荧光生物传感器方面具有相当大的前景，但它们遇到了某些困难，例如灵敏度不足、过早激活、需要外源推进力以及反应速率低。在本次贡献中，探索了多种有利可图的解决方案。首先，集成催化发夹组装（CHA）实现的非酶等温核酸扩增以增强灵敏度。随后，一种DNA成分被简单地用光裂解键功能化以进行光响应方式，从而仅当生物传感器暴露于外部紫外光源时才会发生目标识别，从而克服了生物递送期间的过早激活。此外，通过将生物硫醇还原成MnO 2纳米片来实现特殊的自推进行走机制，从而向Mn 2+依赖的DNAzyme自提供推进力。通过携带DNA分子框架的生物传感系统，产生独特的浓度定位效应，使核酸接触反应速率显着提高6倍。随后，癌症相关 microRNA 生物标志物 (miRNA-21) 的超灵敏体外检测性能得到验证，检测限低至 2.89 fM。特别重要的是，我们的多个概念结合的3D DNA步行放大器能够在活细胞甚至身体中实现高效的荧光生物成像，在疾病分析中展现出良好的应用前景。
更新日期：2024-01-23
中文翻译：
光响应 CHA 集成自推进 3D DNA 步行放大器在 DNA 分子框架的浓度定位效应内实现高效荧光生物成像
虽然三维 (3D) DNA 步行放大器在构建用于生物成像的先进的基于 DNA 的荧光生物传感器方面具有相当大的前景，但它们遇到了某些困难，例如灵敏度不足、过早激活、需要外源推进力以及反应速率低。在本次贡献中，探索了多种有利可图的解决方案。首先，集成催化发夹组装（CHA）实现的非酶等温核酸扩增以增强灵敏度。随后，一种DNA成分被简单地用光裂解键功能化以进行光响应方式，从而仅当生物传感器暴露于外部紫外光源时才会发生目标识别，从而克服了生物递送期间的过早激活。此外，通过将生物硫醇还原成MnO 2纳米片来实现特殊的自推进行走机制，从而向Mn 2+依赖的DNAzyme自提供推进力。通过携带DNA分子框架的生物传感系统，产生独特的浓度定位效应，使核酸接触反应速率显着提高6倍。随后，癌症相关 microRNA 生物标志物 (miRNA-21) 的超灵敏体外检测性能得到验证，检测限低至 2.89 fM。特别重要的是，我们的多个概念结合的3D DNA步行放大器能够在活细胞甚至身体中实现高效的荧光生物成像，在疾病分析中展现出良好的应用前景。
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