Download Development of luminescent probes for selective detection of double

DEVELOPMENT OF LUMINESCENT PROBES FOR SELECTIVE DETECTION OF
DOUBLE STRANDED DNA
Mykhaylo Yu. Losytskyy, Vladyslava B. Kovalska, Sergiy S. Lukashov,
& Sergiy M. Yarmoluk
Institute of Molecular Biology and Genetics, NASci of Ukraine, 150 Zabolotnogo St.,
03143 Kyiv, Ukraine; Теl/fax: 380 44 522 24 58
[email protected]
Development of fluorescent probes that selectively interact with double stranded (ds) DNA is actual for
several modern diagnostics and research methods. For today cyanine dyes are the most sensitive
fluorescent probes for nucleic acids detection. But majority of widely used cyanines demonstrate
comparable sensitivity both to dsDNA and single stranded DNA or RNA. Earlier we proposed trimethine
cyanine dyes with crescent molecule shape as efficient groove binders, interacting with dsDNA with up
to hundred times emission increasing and demonstrating high selectivity to dsDNA as compared with
RNA [1].
For representative of family of trimethine cyanines – iso-propyl meso-substituted benzothiazole dye
Cyan βiPr the ability to form luminescent aggregates (J-aggregates) selectively in the presence of dsDNA
was shown [2]. The high preference of Cyan βiPr to formation of the J-aggregates on AT-containing
polynucleotides as compared to GC-containing ones points on the formation of the aggregates in the
dsDNA groove. Both absorption and fluorescence spectra maxima of such J-aggregates are shifted as
compared to the corresponding spectra of the non-aggregated dye (Table). Applicability of Cyan βiPr for
single molecule detection techniques was shown by the studies of the dye in presence of poly(dA)poly(dT) polynucleotide by fluorescent microscopy. This way red fluorescent images corresponding to
the emission of dye J-aggregates were obtained.
Table. Spectral-luminescent characteristics of Cyan βiPr dye.
free dye
in DNA presence
ex, nm
em, nm
ex, nm
em, nm
IDNA/Ifree
554
571
553 / 595*
572 / 605*
16.7 / 20.7*
ex(em) - excitation (emission)spectrum maximum, IDNA/Ifree – dye emission intensity increasing upon
binding with DNA; * – characteristics of J-aggregate band.
Thus we propose meso-substituted trimethine cyanine dyes as probes for using in techniques that require
selective detection of dsDNA in solution as well as for the visualization of dsDNA molecules in
fluorescent microscopy.
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References:
1. V.B. Kovalska, K.D. Volkova, M.Yu. Losytskyy, O.I. Tolmachev, A.O. Balanda and S.M.
Yarmoluk, 6,6'-Disubstituted benzothiazole trimethine cyanines – new fluorescent dyes for DNA
detection. Spectrochim. Acta Part A 65, 271-277, 2006.
2. M.Yu. Losytskyy, V.M. Yashchuk, S.S. Lukashov, S.M. Yarmoluk. Davydov Splitting in Spectra of
Cyanine Dye J-Aggregates, Formed on the Polynucleotide. Journal of Fluorescence 12, 109-112,
2002.
Acknowledgements: The work was partially supported by STCU-NASU Project # 4936.
www.isranalytica.org.il
Organized and Produced by:
P.O.B 4034 Ness-Ziona 70400, Israel
Tel: +972-8-931-3070, Fax: +972-8-931-3071
Site: www.BioForum.org.il
E-mail: [email protected]