分析上转型 RNA 适体的荧光性质
Characterizing Fluorescence Properties of Turn-on RNA Aptamers.
机构信息
Biochemistry and Biophysics Center, National Heart, Lung and Blood Institute, Bethesda, MD, USA.
出版信息
Methods Mol Biol. 2023;2568:25-36. doi: 10.1007/978-1-0716-2687-0_3.
Abstract
Fluorescent RNA aptamers are tools for studying RNA localization and interactions in vivo. The photophysical properties of these in vitro selected RNAs should be characterized prior to cellular imaging experiments. Here, we describe the process of determining the fluorophore affinity, fluorescence enhancement, and fluorescence lifetime(s) of the Mango-III fluorescence turn-on aptamer. Parameters determined through these protocols will aid in establishing conditions for live-cell imaging.
摘要
荧光 RNA 适体是研究体内 RNA 定位和相互作用的工具。在进行细胞成像实验之前，应对这些体外选择的 RNA 的光物理性质进行表征。本文描述了测定芒果 III 荧光开启适体的荧光团亲和力、荧光增强和荧光寿命的过程。通过这些方案确定的参数将有助于建立活细胞成像的条件。
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