基于功能性核酸识别的荧光各向异性分析方法与应用

荧光各向异性方法又称荧光偏振法,基于相互作用的分子结合前后发射光退偏振的不同而实现对相互作用的研究或对目标物的检测。20世纪50年代Gregorio Weber用荧光各向异性法研究了氮磺酰氯与牛血清白蛋白和卵清白蛋白的作用,开启了该方法在生物化学研究中应用的先河。而20世纪90年代开始的功能性核酸(FNAs,包括核酸适配体和核酸酶等)的发现与合成,使基于功能性核酸的传感得到了广泛的应用。核酸适配体能够特异性识别目标分子,基于核酸识别的荧光各向异性分析方法具有高选择性、高灵敏度、高通量等优势,在研究蛋白质、核酸和小分子的相互作用中起到重要作用,然而如何提高结合前后的荧光各向异性信号变化,尤其是小分子识别前后,在基于功能性核酸识别的分析方法发展中是一个挑战。介绍了基于功能性核酸识别的荧光各向异性的方法应用于检测蛋白质、核酸及其他在生命活动中起重要作用的小分子的基本原理及设计理念。

Fluorescence Anisotropy and Applications Based on Functional Nucleic Acid Recognition

Based on the difference of the depolarization of the emitted light before and after the interaction of molecules,fluores-cence anisotropy,also known as fluorescence polarization,can be beneficial for the study of interactions and the detection of the targets.In 1950s,Gregorio Weber first studied the interactions between dansyl chloride and bovine serum albumin or ovalbumin with fluorescence anisotropy method,which paved the way for fluorescence anisotropy in biochemical applications.Since the ear-ly 1990s,functional nucleic acids (FNAs,including aptamers,and nucleic acid enzymes)were discovered and synthesized,which have been widely used in functional nucleic acid-based sensing.Aptamers are oligonucleotides that can recognize and bind specif-ically to various molecular targets.The fluorescence anisotropy methods which are based on aptamer recognition have the advan-tages of high selectivity,sensitivity and through-put.They play an important role in the study of interaction with protein,nucle-ic acids and small molecules.However,the way of enhancing the fluorescence anisotropy change of small molecules associated with the binding events is challenging.This paper reviews the basic principles and designs of fluorescence anisotropy methods bases on functional nucleic acid recognition for study and detection of proteins,nucleic acids and small molecules that play an im-portant role in the life activity.