Molecular design of Cy3 derivative for highly sensitive in-stem molecular beacon and its application to the wash-free FISH

被引：15
作者：
Kashida, Hiromu [1 ,2 ]
Osawa, Takuya [1 ]
Morimoto, Kazuhiro [1 ]
Kamiya, Yukiko [1 ,3 ]
Asanuma, Hiroyuki [1 ]
机构：
[1] Nagoya Univ, Grad Sch Engn, Chikusa Ku, Nagoya, Aichi 4648603, Japan
[2] Japan Sci & Technol Agcy, PRESTO, Kawaguchi, Saitama 3320012, Japan
[3] Nagoya Univ, EcoTopia Sci Inst, Chikusa Ku, Nagoya, Aichi 4648603, Japan
来源：
BIOORGANIC & MEDICINAL CHEMISTRY | 2015年 / 23卷 / 08期
关键词：
DNA; Molecular beacon; Cy3; Fluorescence in situ hybridization; RATIOMETRIC BIMOLECULAR BEACONS; LIVING CELLS; BACKGROUND EMISSION; RNA DETECTION; DNA; HYBRIDIZATION; PROBES; FLUORESCENCE; EXPRESSION;
D O I：
10.1016/j.bmc.2015.02.030
中图分类号：
Q5 [生物化学]; Q7 [分子生物学];
学科分类号：
071010 ; 081704 ;
摘要：
We herein describe a novel in-stem molecular beacon (ISMB) containing multiple Cy3-quencher pairs on D-threoninol scaffolds in the stem region. The designed Cy3 derivative was not significantly quenched by the adjacent nucleobases, self-quenching of the fluorophore was minimal, and the fluorophore did not severely destabilize the duplex. Using newly designed Cy3, we synthesized ISMBs containing two Cy3 moieties. The signal to background ratio of the ISMB containing two Cy3 moieties was above 100, whereas that with one Cy3 was 30. A Cy3-derivative containing ISMB used in a fluorescence in situ hybridization (FISH) detected endogenous beta-actin mRNA in fixed cells without need for washing procedures. (C) 2015 Elsevier Ltd. All rights reserved.
引用
收藏
页码：1758 / 1762
页数：5
相关论文
共 32 条
[1]   Quencher-free linear probe with multiple fluorophores on an acyclic scaffold [J].
Asanuma, Hiroyuki ;
Akahane, Mariko ;
Kondo, Nobuyo ;
Osawa, Takuya ;
Kato, Tomohiro ;
Kashida, Hiromu .
CHEMICAL SCIENCE, 2012, 3 (11) :3165-3169
[2]   A polycation-chaperoned in-stem molecular beacon system [J].
Asanuma, Hiroyuki ;
Osawa, Takuya ;
Kashida, Hiromu ;
Fujii, Taiga ;
Liang, Xingguo ;
Niwa, Kosuke ;
Yoshida, Yasuko ;
Shimadad, Naohiko ;
Maruyama, Atsushi .
CHEMICAL COMMUNICATIONS, 2012, 48 (12) :1760-1762
[3]   The nature of the different environmental sensitivity of symmetrical and unsymmetrical cyanine dyes: an experimental and theoretical study [J].
Cao, Jianfang ;
Wu, Tong ;
Hu, Chong ;
Liu, Tao ;
Sun, Wen ;
Fan, Jiangli ;
Peng, Xiaojun .
PHYSICAL CHEMISTRY CHEMICAL PHYSICS, 2012, 14 (39) :13702-13708
[4]   Ratiometric bimolecular beacons for the sensitive detection of RNA in single living cells [J].
Chen, Antony K. ;
Davydenko, Olga ;
Behlke, Mark A. ;
Tsourkas, Andrew .
NUCLEIC ACIDS RESEARCH, 2010, 38 (14) :e148-e148
[5]   Molecular beacons with a homo-DNA stem: improving target selectivity [J].
Crey-Desbiolles, C ;
Ahn, DR ;
Leumann, CJ .
NUCLEIC ACIDS RESEARCH, 2005, 33 (08) :1-7
[6]   Single-mismatch detection using gold-quenched fluorescent oligonucleotides [J].
Dubertret, B ;
Calame, M ;
Libchaber, AJ .
NATURE BIOTECHNOLOGY, 2001, 19 (04) :365-370
[7]   Bulge-like Asymmetric Heterodye Clustering in DNA Duplex Results in Efficient Quenching of Background Emission Based on the Maximized Excitonic Interaction [J].
Fujii, Taiga ;
Hara, Yuichi ;
Osawa, Takuya ;
Kashida, Hiromu ;
Liang, Xingguo ;
Yoshida, Yasuko ;
Asanuma, Hiroyuki .
CHEMISTRY-A EUROPEAN JOURNAL, 2012, 18 (35) :10865-10872
[8]   A Highly Sensitive, Excimer-Controlled Molecular Beacon [J].
Haener, Robert ;
Biner, Sarah M. ;
Langenegger, Simon M. ;
Meng, Tao ;
Malinovskii, Vladimir L. .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2010, 49 (07) :1227-1230
[9]   Coherent Quenching of a Fluorophore for the Design of a Highly Sensitive In-Stem Molecular Beacon [J].
Hara, Yuichi ;
Fujii, Taiga ;
Kashida, Hiromu ;
Sekiguchi, Koji ;
Liang, Xingguo ;
Niwa, Kosuke ;
Takase, Tomokazu ;
Yoshida, Yasuko ;
Asanuma, Hiroyuki .
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION, 2010, 49 (32) :5502-5506
[10]   Brightness Enhanced DNA FIT-Probes for Wash-Free RNA Imaging in Tissue [J].
Hoevelmann, Felix ;
Gaspar, Imre ;
Ephrussi, Anne ;
Seitz, Oliver .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2013, 135 (50) :19025-19032
1234