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基于粒子内滚环扩增的核-卫星型磁性超结构用于 microRNA 检测。

On-Particle Rolling Circle Amplification-Based Core-Satellite Magnetic Superstructures for MicroRNA Detection.

机构信息

Department of Engineering Sciences, Uppsala University, The Ångström Laboratory , Box 534, SE-751 21 Uppsala, Sweden.

Department of Immunology, Genetics and Pathology, Uppsala University, The Rudbeck Laboratory , SE-751 85 Uppsala, Sweden.

出版信息

ACS Appl Mater Interfaces. 2018 Jan 24;10(3):2957-2964. doi: 10.1021/acsami.7b16293. Epub 2018 Jan 8.

Abstract

Benefiting from the specially tailored properties of the building blocks as well as of the scaffolds, DNA-assembled core-satellite superstructures have gained increasing interest in drug delivery, imaging, and biosensing. The load of satellites plays a vital role in core-satellite superstructures, and it determines the signal intensity in response to a biological/physical stimulation/actuation. Herein, for the first time, we utilize on-particle rolling circle amplification (RCA) to prepare rapidly responsive core-satellite magnetic superstructures with a high load of magnetic nanoparticle (MNP) satellites. Combined with duplex-specific nuclease-assisted target recycling, the proposed magnetic superstructures hold great promise in sensitive and rapid microRNA detection. The long single-stranded DNA produced by RCA serving as the scaffold of the core-satellite superstructure can be hydrolyzed by duplex-specific nuclease in the presence of target microRNA, resulting in a release of MNPs that can be quantified in an optomagnetic sensor. The proposed biosensor has a simple mix-separate-measure strategy. For let-7b detection, the proposed biosensor offers a wide linear detection range of approximately 5 orders of magnitude with a detection sensitivity of 1 fM. Moreover, it has the capability to discriminate single-nucleotide mismatches and to detect let-7b in cell extracts and serum, thus showing considerable potential for clinical applications.

摘要

得益于构建基块和支架的特殊性质,DNA 组装的核-卫星超结构在药物输送、成像和生物传感方面引起了越来越多的关注。卫星的负载在核-卫星超结构中起着至关重要的作用,它决定了对生物/物理刺激/触发的信号强度。在此,我们首次利用颗粒上滚环扩增(RCA)来快速制备具有高负载磁性纳米颗粒(MNP)卫星的快速响应核-卫星磁性超结构。结合双链特异性核酸酶辅助的靶标循环回收,所提出的磁性超结构在敏感和快速 microRNA 检测方面具有很大的应用潜力。RCA 产生的长单链 DNA 作为核-卫星超结构的支架,在存在靶标 microRNA 的情况下,双链特异性核酸酶可以将其水解,从而释放出可以在光磁传感器中定量的 MNPs。所提出的生物传感器具有简单的混合-分离-测量策略。对于 let-7b 的检测,所提出的生物传感器提供了大约 5 个数量级的宽线性检测范围,检测灵敏度为 1 fM。此外,它还具有区分单碱基错配和检测细胞提取物和血清中 let-7b 的能力,因此在临床应用中具有很大的潜力。

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