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Fluorescent properties of oligonucleotide-conjugated thiazole orange probes

Fluorescent properties of oligonucleotide-conjugated thiazole orange probes
Fluorescent properties of oligonucleotide-conjugated thiazole orange probes
The fluorescence properties of thiazole orange, linked via a (1) hydrophobic alkyl or a (2) hydrophillic ethylene glycol chain to the central internucleotidic phosphate group of a pentadeca-2'-deoxyriboadenylate (dA(15)), are evaluated. Linkage at the phosphate group yields two stereoisomers, S-isomer of the phosphorus chiral center (Sp) and R-isomer of the phosphorus chiral center (Rp); these are studied separately. The character of the linkage chain and the chirality of the internucleotidic phosphate linkage site influence the fluorescent properties of these thiazole orange-oligonucleotide conjugates (TO-probes). Quantum yields of fluorescence (Phi(fl)) of between 0.04 and 0.07 were determined for the single-stranded conjugates. The fluorescence yield increased by up to five times upon hybridization with the complementary sequence (d5'[CACT(15)CAC(3')]); Phi(fl) values of between 0.06-0.35 were determined for the double-stranded conjugates. The Phi(fl) value (0.17) of thiazole orange, 1-(N,N'-trimethylaminopropyl)-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiazole)- 2-methylidene]-quinolinium iodide (TO-Pro 1) in the presence of the oligonucleotide duplex (TO-Pro 1: dA(15)(.)d(5')[CACT(15)CAC(3')] (1:1)) is much less than that for some of the hybrids of the conjugates. Our studies, using steady-state and time-resolved fluorescence experiments, show that a number of discrete fluorescent association species between the thiazole orange and the helix are formed. Time-resolved studies on the four double-stranded TO-probes revealed that the fluorescent oligonucleotide-thiazole orange complexes are common, only the distribution of the species varies with the character of the chain and the chirality at the internucleotidic phosphate site. Those TO-probes in which the isomeric structure of the phosphate-chain linkage is Rp, and therefore such that the fluorophore is directed toward the minor groove, have higher Phi(fl) values than the Sp isomer. Of the systems studied, thiazole orange linked by an alkyl chain to the internucleotidic phosphate (Rp isomer) has the highest Phi(fl) and the greatest fraction of the longest-lived fluorescent thiazole orange species (in the hybrid form).
0031-8655
201-210
Privat, E.
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Melvin, T.
fd87f5eb-2bb9-48fa-b7be-7100ace9c50f
Mérola, F.
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Schweizer, G.
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Prodhomme, S.
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Asseline, U.
5f76eeb4-9779-4967-95ee-2c2b410f26fe
Vigny, P.
18506220-1c8e-4970-a067-d3f9326f4ecb
Privat, E.
4d1070de-c9f6-491c-98ca-1d1cc077eb45
Melvin, T.
fd87f5eb-2bb9-48fa-b7be-7100ace9c50f
Mérola, F.
e8fe3b1b-094e-450d-8938-d9ad24c343ba
Schweizer, G.
91f6275a-1323-48a1-8af7-1e9d1563b76e
Prodhomme, S.
944412fa-2548-4b97-8c49-699bf286590b
Asseline, U.
5f76eeb4-9779-4967-95ee-2c2b410f26fe
Vigny, P.
18506220-1c8e-4970-a067-d3f9326f4ecb

Privat, E., Melvin, T., Mérola, F., Schweizer, G., Prodhomme, S., Asseline, U. and Vigny, P. (2002) Fluorescent properties of oligonucleotide-conjugated thiazole orange probes. Photochemistry and Photobiology, 75 (3), 201-210. (doi:10.1562/0031-8655(2002)0750201FPOOCT2.0.CO2).

Record type: Article

Abstract

The fluorescence properties of thiazole orange, linked via a (1) hydrophobic alkyl or a (2) hydrophillic ethylene glycol chain to the central internucleotidic phosphate group of a pentadeca-2'-deoxyriboadenylate (dA(15)), are evaluated. Linkage at the phosphate group yields two stereoisomers, S-isomer of the phosphorus chiral center (Sp) and R-isomer of the phosphorus chiral center (Rp); these are studied separately. The character of the linkage chain and the chirality of the internucleotidic phosphate linkage site influence the fluorescent properties of these thiazole orange-oligonucleotide conjugates (TO-probes). Quantum yields of fluorescence (Phi(fl)) of between 0.04 and 0.07 were determined for the single-stranded conjugates. The fluorescence yield increased by up to five times upon hybridization with the complementary sequence (d5'[CACT(15)CAC(3')]); Phi(fl) values of between 0.06-0.35 were determined for the double-stranded conjugates. The Phi(fl) value (0.17) of thiazole orange, 1-(N,N'-trimethylaminopropyl)-4-[3-methyl-2,3-dihydro-(benzo-1,3-thiazole)- 2-methylidene]-quinolinium iodide (TO-Pro 1) in the presence of the oligonucleotide duplex (TO-Pro 1: dA(15)(.)d(5')[CACT(15)CAC(3')] (1:1)) is much less than that for some of the hybrids of the conjugates. Our studies, using steady-state and time-resolved fluorescence experiments, show that a number of discrete fluorescent association species between the thiazole orange and the helix are formed. Time-resolved studies on the four double-stranded TO-probes revealed that the fluorescent oligonucleotide-thiazole orange complexes are common, only the distribution of the species varies with the character of the chain and the chirality at the internucleotidic phosphate site. Those TO-probes in which the isomeric structure of the phosphate-chain linkage is Rp, and therefore such that the fluorophore is directed toward the minor groove, have higher Phi(fl) values than the Sp isomer. Of the systems studied, thiazole orange linked by an alkyl chain to the internucleotidic phosphate (Rp isomer) has the highest Phi(fl) and the greatest fraction of the longest-lived fluorescent thiazole orange species (in the hybrid form).

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Published date: March 2002

Identifiers

Local EPrints ID: 49830
URI: http://eprints.soton.ac.uk/id/eprint/49830
ISSN: 0031-8655
PURE UUID: cb474e90-12c6-488e-a917-35c48da6a4ef

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Date deposited: 03 Dec 2007
Last modified: 13 Mar 2019 20:53

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Contributors

Author: E. Privat
Author: T. Melvin
Author: F. Mérola
Author: G. Schweizer
Author: S. Prodhomme
Author: U. Asseline
Author: P. Vigny

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