Large-Stokes-shift-based folded DNA probing systems targeting DNA and miRNA 21 with signal amplification.

Autor: Le BH; Department of Bioactive Material Sciences, Chonbuk National University, Jeonju, Jeonbuk 561-567, South Korea., Nguyen TT; Department of Bioactive Material Sciences, Chonbuk National University, Jeonju, Jeonbuk 561-567, South Korea., Joo HN; Department of Chemistry, Chonbuk National University, Jeonju, Jeonbuk 561-567, South Korea., Seo YJ; Department of Bioactive Material Sciences, Chonbuk National University, Jeonju, Jeonbuk 561-567, South Korea; Department of Chemistry, Chonbuk National University, Jeonju, Jeonbuk 561-567, South Korea. Electronic address: yseo@jbnu.ac.kr.
Jazyk: angličtina
Zdroj: Bioorganic & medicinal chemistry [Bioorg Med Chem] 2018 Sep 15; Vol. 26 (17), pp. 4881-4885. Date of Electronic Publication: 2018 Aug 25.
DOI: 10.1016/j.bmc.2018.08.027
Abstrakt: Large-Stokes-shift based simple folded DNA probing system (LSFP) had a simple folded DNA structure and exhibited a large Stokes-shifted (194 nm) fluorescence signal upon excitation at a single wavelength (386 nm). This Stokes shift was achieved through a simple combination of donor and acceptor fluorophores and employing multi-FRET systematically. This unique large Stokes-shifted fluorescence signal was used to detect target DNA with large increases in the fluorescence signal (9.7-14.2 fold). This LSFP exhibited enough selectivity, distinguishing a perfectly matched sequence from the probe itself and mismatched sequences. Surprisingly, when DSN was used for signal amplification with miR21P probing system whose target is miRNA 21, it showed high sensitivity (13.7 aM) and selectivity (one base mismatch discrimination). This system has several advantages over other molecular beacons (MBs): (i) it is easy to design and synthesize the probing system that does not require the construction of a finely designed stem and loop, as in most MBs (this can prevent the degradation of miR21P itself by DSN enzyme without special backbone modification); (ii) it can control unique fluorescence, such as large Stokes-shifted fluorescence through a simple combination of donor and acceptor fluorophores; and (iii) through signal amplification using DSN, it can efficiently detect extremely small amounts of target miRNA with high sensitivity (13.7 aM).
(Copyright © 2018 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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