Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON, Canada.
Department of Chemistry, Waterloo Institute for Nanotechnology, University of Waterloo, Waterloo, ON, Canada.
Methods Enzymol. 2021;651:373-396. doi: 10.1016/bs.mie.2021.01.013. Epub 2021 Mar 11.
Highly sensitive and selective detection of lanthanide ions is a major analytical challenge. In recent years, the use of DNA for this purpose has been pursued. For such highly charged cations, it is difficult to select their aptamers due to strong nonspecific binding. On the other hand, the use of catalytic DNA or DNAzymes has an advantage to overcome this problem, especially DNAzymes with RNA-cleaving activity. In this chapter, a few such DNAzymes are introduced and methods for in vitro selection of lanthanide-dependent RNA-cleaving DNAzymes are described in detail, including the selection protocols, the DNA sequences used, the characterization of selected DNAzymes and their conversion into biosensors. All of the experiments use only fluorophore-labeled DNA, and radioisotope labeling is completely avoided. The resulting DNAzymes can distinguish lanthanides from non-lanthanide metals, tell the difference between light and heavy lanthanides, and can be used together to discriminate individual lanthanides.
高灵敏度和选择性地检测镧系元素离子是一个主要的分析挑战。近年来，人们一直在探索使用 DNA 来实现这一目标。对于这种带高电荷的阳离子，由于强非特异性结合，很难选择它们的适体。另一方面，使用催化 DNA 或 DNA 酶具有克服这一问题的优势，特别是具有 RNA 切割活性的 DNA 酶。在本章中，介绍了几种这样的 DNA 酶，并详细描述了体外选择依赖镧系元素的 RNA 切割 DNA 酶的方法，包括选择方案、使用的 DNA 序列、所选 DNA 酶的特性及其转化为生物传感器。所有的实验都只使用荧光标记的 DNA，完全避免了放射性同位素标记。所得的 DNA 酶可以区分镧系元素和非镧系金属，区分轻镧系元素和重镧系元素，并且可以一起用于区分个别镧系元素。
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