Synthesis, serum stability and cell uptake of cyclic and hairpin decoy Oligonucleotides for TCF/LEF and GLI transcription factors
Synthesis, serum stability and cell uptake of cyclic and hairpin decoy Oligonucleotides for TCF/LEF and GLI transcription factors
A series of hairpin oligonucleotides bearing a 5?-alkyne and 3?-azide were cyclised using the CuAAC reaction to form cyclic decoys for targeting the DNA binding site of the TCF/LEF and GLI transcription factors. Incubation of a fluorescent hairpin oligomer and its cyclic analogue in fetal calf serum showed that the cyclic construct has significantly greater stability to enzymatic degradation. Cell uptake studies using HEK-293 cells with the fluorescent cyclic decoy in the presence of lipofectamine 2000 transfection agent indicated that this analogue is taken up by the cells and localizes to the cell nucleus. Localized fluorescence was observed in the nuclei of HEK-293 cells after only 1.5 h incubation which increased over a period of 4 h and persisted for 24 h.
cyclic oligonucleotide, decoy, click chemistry, serum stability, cell uptake
367-372
El-Sagheer, Afaf H.
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
December 2008
El-Sagheer, Afaf H.
05b8295a-64ad-4fdf-ad57-c34934a46c04
Brown, Tom
a64aae36-bb30-42df-88a2-11be394e8c89
El-Sagheer, Afaf H. and Brown, Tom
(2008)
Synthesis, serum stability and cell uptake of cyclic and hairpin decoy Oligonucleotides for TCF/LEF and GLI transcription factors.
International Journal of Peptide Research and Therapeutics, 14 (4), .
(doi:10.1007/s10989-008-9153-0).
Abstract
A series of hairpin oligonucleotides bearing a 5?-alkyne and 3?-azide were cyclised using the CuAAC reaction to form cyclic decoys for targeting the DNA binding site of the TCF/LEF and GLI transcription factors. Incubation of a fluorescent hairpin oligomer and its cyclic analogue in fetal calf serum showed that the cyclic construct has significantly greater stability to enzymatic degradation. Cell uptake studies using HEK-293 cells with the fluorescent cyclic decoy in the presence of lipofectamine 2000 transfection agent indicated that this analogue is taken up by the cells and localizes to the cell nucleus. Localized fluorescence was observed in the nuclei of HEK-293 cells after only 1.5 h incubation which increased over a period of 4 h and persisted for 24 h.
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Published date: December 2008
Keywords:
cyclic oligonucleotide, decoy, click chemistry, serum stability, cell uptake
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Local EPrints ID: 146697
URI: http://eprints.soton.ac.uk/id/eprint/146697
ISSN: 1573-3149
PURE UUID: e72ff831-3ec4-4984-b4a3-793e4a707cfc
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Date deposited: 22 Apr 2010 08:58
Last modified: 14 Mar 2024 02:51
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Author:
Afaf H. El-Sagheer
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