Two Successive Reactions on a DNA Template: A Strategy for Improving Background Fluorescence and Specificity in Nucleic Acid Detection

被引：33

作者：

Franzini, Raphael M. ^{[1
]}

Kool, Erie T. ^{[1
]}

机构：

[1] Stanford Univ, Dept Chem, Stanford, CA 94305 USA

来源：

CHEMISTRY-A EUROPEAN JOURNAL | 2011年 / 17卷 / 07期

基金：

美国国家卫生研究院;

关键词：

fluorescence; nucleic acids; quenched probes; Staudinger reaction; template synthesis; PROBE ACTIVATION; MESSENGER-RNA; BINARY PROBES; CELLS; HYBRIDIZATION; REDUCTION; DISCRIMINATION; AMPLIFICATION; AUTOLIGATION; CHEMISTRY;

D O I：

10.1002/chem.201002426

中图分类号：

O6 [化学];

学科分类号：

070301 [无机化学];

摘要：

We report a new strategy for template-mediated fluorogenic chemistry that results in enhanced performance for the fluorescence detection of nucleic acids. In this approach, two successive templated reactions are required to induce a fluorescence signal, rather than only one. These novel fluorescein-labeled oligonucleotide probes, termed 2-STAR (STAR=Staudinger-triggered alpha-azidoether release) probes, contain two quencher groups tethered by separate reductively cleavable linkers. When a 2-STAR quenched probe successively binds adjacent to two mono-triphenylphosphine-(TPP)-DNAs or one dual-TPP-DNA, the two quenchers are released, resulting in a fluorescence signal. Because of the requirement for two consecutive reactions, 2-STAR probes display an unprecedented level of sequence specificity for template-mediated probe designs. At the same time, background emission generated by off-template reactions or incomplete quenching is among the lowest of any fluorogenic reactive probes for the detection of DNA or RNA.

引用

页码：2168 / 2175

页数：8

共 46 条

[1]

Flow cytometric detection of specific RNAs in native human cells with quenched autoligating FRET probes [J].