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1.
Clustered regularly interspaced short palindromic repeats (CRISPR) is a promising technology in the biological world. As one of the CRISPR-associated (Cas) proteins, Cas12a is an RNA-guided nuclease in the type V CRISPR-Cas system, which has been a robust tool for gene editing. In addition, due to the discovery of target-binding-induced indiscriminate single-stranded DNase activity of Cas12a, CRISPR-Cas12a also exhibits great promise in biosensing. This minireview not only gives a brief introduction to the mechanism of CRISPR-Cas12a but also highlights the recent developments and applications in biosensing and gene regulation. Finally, future prospects of the CRISPR-Cas12a system are also discussed. We expect this minireview will inspire innovative work on the CRISPR-Cas12a system by making full use of its features and advantages.  相似文献   
2.
CRISPR-Cas12a系统的反式切割活性在其识别特定的DNA激活序列后被激活,这不仅能实现特定DNA靶标的直接定量分析,同时也为构建针对多种生物标志物的体外传感体系带来了新的思路。然而,已有文献中所采用的双链DNA(dsDNA)和单链DNA(ssDNA)激活序列结构多种多样,缺乏全面、系统的设计指导原则。针对该问题,该文系统研究了不同结构的DNA激活序列对LbaCas12a反式切割活性的影响。通过对比研究,得出以下结论:(1)前间区序列邻近基序(PAM)位点有助于LbaCas12a更高效地靶向结合dsDNA激活序列和ssDNA激活序列;(2)PAM近端区域缺少序列片段会降低Cas12a-crRNA定位激活序列的效率;(3)删除PAM远端序列片段有利于增强LbaCas12a的反式切割活性;(4)由于省略了dsDNA解链过程,ssDNA激活序列在激活LbaCas12a的反式切割活性方面普遍比dsDNA激活序列产生的效果更好。根据这些发现,该文提出了一种LbaCas12a所青睐的高效激活序列结构,其激活的LbaCas2a反式酶切活性较采用含PAM位点的标准dsDNA激活序列高出3.7倍。研究结果为构建基于CRISPR-Cas12a的高效体外生物传感系统提供了重要支撑。  相似文献   
3.
The synthetic estrogen 17α‐ethinylestradiol (EE2) is an active component of oral contraceptives. It is considered as an endocrine disrupting compound that, once incorporated into an organism, affects the hormonal balance of animals and humans. In this study we characterized the DNA‐EE2 interaction using an electrochemical biosensor and biosensing in solution phase with the double stranded DNA from salmon sperm and deoxyguanosine monophosphate (dGMP). Differential pulse voltammetry method has been applied based on voltammetric anodic responses of the deoxyguanine (dGuo) and deoxyadenine (dAdo) as well as EE2 in the medium of phosphate buffer solution pH 7.0. Binding of EE2 to the nucleobases leads to a decrease of their anodic signals. Association constant for DNA‐EE2 interaction has been estimated to be about 1.1 ? 103 L mol?1 and 1.4 ? 103 L mol?1 for dGuo and dAdo responses, respectively. The association is reversible as indicated by decrease of the EE2 response in pure buffer solution due to leaching of EE2 from the surface attached DNA. The DNA‐EE2 association has been confirmed also by UV‐vis spectrometric experiments.  相似文献   
4.
刘超  田飞  邓瑾琦  孙佳姝 《化学学报》2022,80(5):679-689
复杂生命体系中关键分子及微纳生物粒子的高灵敏、高特异检测, 对理解多层次多尺度生物学过程、阐明疾病发生发展机制和探索新型生物标志物等具有重要意义. 微流控生物传感器整合了微流控技术和生物传感技术的诸多优势, 在微量生物样本精准测量方面取得了显著进展. 近年来, 微流控热泳生物传感技术(Thermomicrofluidic biosensing)利用物质在局域温度梯度场中的热泳定向迁移现象, 并结合均相生物传感及信号放大新策略, 实现了复杂样本中生物分子及微纳生物粒子的快速、高灵敏、原位检测. 重点阐述了以热泳为核心的微流控传感技术, 包括微量热泳、热泳-对流耦合、热泳-扩散泳耦合以及热泳-电泳耦合等方法, 总结了不同传感方法的原理、特点及其在生物分子(蛋白、核酸等)与微纳生物粒子(细胞外囊泡、病毒、细胞等)检测中的应用, 并探讨了微流控热泳技术在生物医学检测领域中面临的挑战与未来发展方向.  相似文献   
5.
Artificial and natural lipid membranes that elicit transmembrane signaling is are useful as a platform for channel‐based biosensing. In this account we summarize our research on the design of transmembrane signaling associated with lipid bilayer membranes containing nanopore‐forming compounds. Channel‐forming compounds, such as receptor ion‐channels, channel‐forming peptides and synthetic channels, are embedded in planar and spherical bilayer lipid membranes to develop highly sensitive and selective biosensing methods for a variety of analytes. The membrane‐bound receptor approach is useful for introducing receptor sites on both planar and spherical bilayer lipid membranes. Natural receptors in biomembranes are also used for designing of biosensing methods.  相似文献   
6.
碳量子点是一类具有优异的荧光性能和高生物相容性的纳米材料,在很多领域有广泛的应用,是目前研究的热点材料。本文介绍了碳量子点不同的合成方法,以及碳量子点的荧光、化学发光、电化学发光、类过氧化物酶的活性及毒性等性能的最新研究进展。此外,还对碳量子点在生物传感、生物成像及药物传递等生物医学应用进行了概述。  相似文献   
7.
杨爽  杨贤鹏  王宝俊  王蕾 《化学进展》2021,33(12):2309-2315
纸基生物传感器由于其具有成本低、操作方便、生物可降解、识别元件用量低等优点,近年来受到了广泛的关注。其中,以功能核酸作为识别元件的纸基荧光生物传感器具有较高的灵敏度、瞬时响应以及实时检测等特性,在便携式传感设备方面展现出巨大的潜力。此外,将核酸作为识别元件的纸基无细胞蛋白合成平台,通过条件合成的报告荧光蛋白可实现对病毒、重金属等目标物的特异性检测,具有良好的应用前景。首先,本文介绍了基于核酸的纸基荧光生物传感器的设计,特别是基于核酸的识别元件与纸基材料的结合方式。其次,总结了基于核酸的纸基荧光生物传感器在临床诊断、食品安全检测、环境污染物检测等不同领域的最新研究进展,讨论了其优势与局限性。最后,探讨了基于核酸的纸基荧光生物传感器的发展方向与应用前景,以期为相关领域的研究提供参考。  相似文献   
8.
《Electroanalysis》2017,29(5):1310-1315
A novel photo‐induced electrochemical biosensing method has been developed based on fluorescence quenching effect and electrochemical method. In this sensing strategy, the molecular beacon probes labeled with methylene blue were immobilized on the gold nanoparticles modified gold electrode surface firstly; then dopamine was assembled on the electrode surface through electrostatic interaction with gold nanoparticles. Under the continuous illumination, the fluorescence of the methylene blue was quenched by the gold nanoparticles before hybridization; after hybridization with the complementary DNA, methylene blue was far away from the gold nanoparticles and the fluorescence recovered, and then singlet oxygen was generated in the photosensitive reaction of methylene blue in the presence of dissolved oxygen. Singlet oxygen reacted with dopamine, which resulted in the reduction of concentration of the dopamine on the electrode surface. The current of the dopamine on the electrode was used for the sensing of the conformational change of molecular beacon and hence for the detection of target DNA.  相似文献   
9.
Reflectometric interference spectroscopy (RIfS) is a label-free, time-resolved technique, and suitable for detecting antibody–antigen interaction. This work describes a continuous flow biosensor for C-reactive protein (CRP), involving an effective immobilization method of a monoclonal antibody against CRP (anti-CRP) to achieve highly sensitive RIfS-based detection of CRP. The silicon nitride-coated silicon chip (SiN chip) for the RIfS sensing was first treated with trimethylsilylchloride (TMS), followed by UV-light irradiation to in situ generation of homogeneous silanols on the surface. Following amination by 3-aminopropyltriethoxysilane, carboxymethyldextran (CMD) was grafted, and subsequently, protein A was immobilized to create the oriented anti-CRP surface. The immobilization process of protein A and anti-CRP was monitored with the RIfS system by consecutive injections of an amine coupling reagent, protein A and anti-CRP, respectively, to confirm the progress of each step in real time. The sensitivity was enhanced when all of the processes were adopted, suggesting that the oriented immobilization of anti-CRP via protein A that was coupled with the grafted CMD on the aminated surface of TMS-treated SiN chip. The feasibility of the present sensing system was demonstrated on the detection of CRP, where the silicon-based inexpensive chips and the simple optical setup were employed. It can be applied to other target molecules in various fields of life science as a substitute of surface plasmon resonance-based expensive sensors.  相似文献   
10.
《Analytical letters》2012,45(9):1785-1799
Abstract

Multiwalled carbon nanotubes (MWNTs) were treated with a mixture of concentrated sulfuric and nitric acid to introduce carboxylic acid groups to the nanotubes. Conducting polymer film was prepared by electrochemical polymerization of neutral red (NR). By using a layer‐by‐layer method, homogeneous and stable MWNTs and poly (neutral red) (PNR) multilayer films were alternately assembled on glassy carbon (GC) electrodes. With the introduction of PNR, the MWNTs/PNR multilayer film system showed synergy between the MWNTs and PNR, with a significant improvement of redox activity due to the excellent electron‐transfer ability of carbon nanotubes (CNTs) and PNR. The electropolymerization is advantageous, providing both prolonged long‐term stability and improved catalytic activity of the resulting modified electrodes. The MWNTs/PNR multilayer film modified glassy carbon electrode allows low potential detection of hydrogen peroxide with high sensitivity and fast response time. As compared to MWNTs and PNR‐modified GC electrodes, the magnitude of the amperometric response of the MWNTs/PNR composite‐modified GC electrode is more than three‐fold greater than that of the MWNTs modified GC electrode, and nine‐fold greater than that of the PNR‐modified GC electrode. With the immobilization of glucose oxidase onto the electrode surface using glutaric dialdehyde, a biosensor that responds sensitively to glucose has been constructed. In pH 6.98 phosphate buffer, nearly interference‐free determination of glucose has been realized at ?0.2 V vs. SCE with a linear range from 50 µM to 10 mM and response time <10s. The detection limit was 10 µM glucose (S/N=3).  相似文献   
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