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琼脂糖凝胶基单分子 FRET 标记法评价 siRNA 稳定性及其与血清成分的相互作用。

Evaluation of siRNA Stability and Interaction with Serum Components Using an Agarose Gel-Based Single-Molecule FRET Labeling Method.

机构信息

Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.

Department of Oncology, Odense University Hospital, Odense, Denmark.

出版信息

Methods Mol Biol. 2021;2282:43-56. doi: 10.1007/978-1-0716-1298-9_4.

Abstract

Small interfering RNAs (siRNAs) are RNA molecules with promising therapeutic potential as a result of their selective mRNA cleavage. However, despite recent progress, low stability in the bloodstream is an impediment to successful administration in vivo. Thus, the availability of flexible and rapid methods for studying siRNA stability and vehicles is crucial for future novel siRNA-based therapeutics. Herein, we report a fast Förster resonance energy transfer (FRET) method based on agarose gel electrophoresis to evaluate the stability of siRNA in serum as well as siRNA interaction with serum proteins and enzymes.

摘要

小干扰 RNA(siRNA)是一种具有治疗潜力的 RNA 分子,因其能够选择性地切割 mRNA。然而,尽管最近取得了进展,但在血液中的低稳定性仍是其体内给药成功的障碍。因此,灵活快速的方法对于研究 siRNA 稳定性和载体至关重要,这对于未来基于 siRNA 的治疗方法至关重要。在此,我们报告了一种基于琼脂糖凝胶电泳的快速Förster 共振能量转移(FRET)方法,用于评估 siRNA 在血清中的稳定性以及 siRNA 与血清蛋白和酶的相互作用。

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