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Journal of Biomedical Optics • Open Access

Fluorescent probe for visualizing guanine-quadruplex DNA by fluorescence lifetime imaging microscopy
Author(s): Ting-Yuan Tseng; Cheng-Hao Chien; Jen-Fei Chu; Wei-Chun Huang; Mei-Ying Lin; Cheng-Chung Chang; Ta-Chau Chang

Paper Abstract

The importance of guanine-quadruplex (G4) is not only in protecting the ends of chromosomes for human telomeres but also in regulating gene expression for several gene promoters. However, the existence of G4 structures in living cells is still in debate. A fluorescent probe, 3,6-bis(1-methyl-2-vinylpyridinium) carbazole diiodide (o-BMVC), for differentiating G4 structures from duplexes is characterized. o-BMVC has a large contrast in fluorescence decay time, binding affinity, and fluorescent intensity between G4 structures and duplexes, which makes it a good candidate for probing G4 DNA structures. The fluorescence decay time of o-BMVC upon interaction with G4 structures of telomeric G-rich sequences is ∼2.8  ns and that of interaction with the duplex structure of a calf thymus is ∼1.2  ns . By analyzing its fluorescence decay time and histogram, we were able to detect one G4 out of 1000 duplexes in vitro. Furthermore, by using fluorescence lifetime imaging microscopy, we demonstrated an innovative methodology for visualizing the localization of G4 structures as well as mapping the localization of different G4 structures in living cells.

Paper Details

Date Published: 9 July 2013
PDF: 7 pages
J. Biomed. Opt. 18(10) 101309 doi: 10.1117/1.JBO.18.10.101309
Published in: Journal of Biomedical Optics Volume 18, Issue 10
Show Author Affiliations
Ting-Yuan Tseng, National Yang-Ming Univ. (Taiwan)
Cheng-Hao Chien, Academia Sinica (Taiwan)
Jen-Fei Chu, Academia Sinica (Taiwan)
Wei-Chun Huang, Academia Sinica (Taiwan)
Mei-Ying Lin, Academia Sinica (Taiwan)
Cheng-Chung Chang, National Chung Hsing Univ. (Taiwan)
Ta-Chau Chang, Academia Sinica (Taiwan)

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