Fujimoto Kazuhisa, Inouye Masahiko
Graduate School of Pharmaceutical Sciences, University of Toyama, Toyama City, Japan.
Yakugaku Zasshi. 2008 Nov;128(11):1605-13. doi: 10.1248/yakushi.128.1605.
In this review, we report DNA duplex-based fluorescence probes/sensors using pyrene monomer-excimer switching. The review mainly comprises two topics: 1) excimer-monomer switching molecular beacons (EMS-MB) and 2) monomer-excimer switching sensors based on the structural motif of antibodies. The EMS-MBs have two pyrene fluorophores connected both at 3' and 5' ends of a single-stranded oligonucleotide. Emission switching occurs from excimer to monomer accompanying isoemissive points when the probes hybridized with target DNAs. The isoemissive points indicate the presence of only two fluorescent species, nonhybridized and hybridized probes in the mixtures, and thereby unambiguous detection of the targets is available. The probes can detect target 19-mer DNAs and can discriminate the targets from their single-nucleotide mismatches at 1 nM concentration. Furthermore, the EMS-MBs have been recently applied to kinetic study for RNase H activity by Tan et al. The structures and emission-switching properties of the EMS-MBs encourage us to develop a new class of fluorescent sensors based on the structural motif of antibodies. The sensors consist of three functional regions, benzo-15-crown-5 ether (or per-O-methylated beta-cyclodextrin), DNA, and pyrene as guest-binding, dimerizing, and sensing sites, respectively. The crown- and CD-modified sensors can detect potassium cation and porphyrin derivatives, respectively, by monomer-excimer emission switching.
在本综述中,我们报道了基于芘单体 - 激基缔合物转换的DNA双链荧光探针/传感器。该综述主要包括两个主题:1)激基缔合物 - 单体转换分子信标(EMS - MB)和2)基于抗体结构基序的单体 - 激基缔合物转换传感器。EMS - MB在单链寡核苷酸的3'和5'末端均连接有两个芘荧光团。当探针与靶DNA杂交时,伴随着等发射点,发射从激基缔合物转换为单体。等发射点表明混合物中仅存在两种荧光物质,即未杂交和杂交的探针,从而可以明确检测靶标。这些探针可以检测19聚体DNA靶标,并能在1 nM浓度下将靶标与其单核苷酸错配区分开来。此外,Tan等人最近已将EMS - MB应用于RNase H活性的动力学研究。EMS - MB的结构和发射转换特性促使我们基于抗体的结构基序开发一类新型荧光传感器。这些传感器由三个功能区域组成,分别为苯并 - 15 - 冠 - 5醚(或全O - 甲基化β - 环糊精)、DNA和芘,作为客体结合、二聚化和传感位点。冠醚和环糊精修饰的传感器可分别通过单体 - 激基缔合物发射转换检测钾阳离子和卟啉衍生物。