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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References
Chan TR, Hilgraf R, Sharpless KB, Fokin VV (2004) Polytriazoles as copper(I)-stabilizing ligands in catalysis. Org Lett 6:2853–2855
Conte MR, Mayol L, Montesarchio D, Piccialli G, Santacroce C (1993) Automated synthesis of cyclic oligodeoxyribonucleotides via phosphoramidite method. Nucleosides Nucleotides 12:351–358
Denapoli L, Galeone A, Mayol L, Messere A, Piccialli G, Santacroce C (1993) Facile preparation of cyclic oligoribonucleotides. J Chem Soc-Perkin Trans 1:747–749
di Magliano MP, Hebrok M (2003) Hedgehog signalling in cancer formation and maintenance. Nat Rev Cancer 3:903–911
Edupuganti OP, Defrancq E, Dumy P (2003) Head-to-tail oxime cyclization of oligodeoxynucleotides for the efficient synthesis of circular DNA analogues. J Org Chem 68:8708–8710
El-Sagheer AH, Kumar R, Findlow S, Werner JM, Lane AN, Brown T (2008) A very stable cyclic DNA mini-duplex with just two base pairs. Chembiochem 9:50–52
Giese K, Amsterdam A, Grosschedl R (1991) DNA-binding properties of the Hmg domain of the lymphoid-specific transcriptional regulator Lef-1. Genes Dev 5:2567–2578
Giusto DA, Knox SM, Lai Y, Tyrelle GD, Aung MT, King GC (2006) Multitasking by multivalent circular DNA aptamers. Chembiochem 7:535–544
Grimau MG, Avino A, Gargallo R, Eritja R (2005) Synthesis and triplex-forming properties of cyclic oligonucleotides with (G,A)-antiparallel strands. Chem Biodivers 2:275–285
Hartig JS, Kool ET (2004) Small circular DNAs for synthesis of the human telomere repeat: varied sizes, structures and telomere-encoding activities. Nucleic Acids Res 32:e152
Ingham PW, McMahon AP (2001) Hedgehog signaling in animal development: paradigms and principles. Genes Dev 15:3059–3087
Katoh M, Katoh M (2007) WNT signaling pathway and stem cell signaling network. Clin Cancer Res 13:4042–4045
Kool ET (1996) Circular oligonucleotides: new concepts in oligonucleotide design. Annu Rev Biophys Biomol Struct 25:1–28
Kumar R, El-Sagheer A, Tumpane J, Lincoln P, Wilhelmsson LM, Brown T (2007) Template-directed oligonucleotide strand ligation, covalent intramolecular DNA circularization and catenation using click chemistry. J Am Chem Soc 129:6859–6864
Lee IK, Ahn JD, Kim HS, Park JY, Lee KU (2003) Advantages of the circular dumbbell decoy in gene therapy and studies of gene regulation. Curr Drug Targets 4:619–623
Polakis P (2000) Wnt signaling and cancer. Genes Dev 14:1837–1851
Rubin LL, de Sauvage FJ (2006) Targeting the Hedgehog pathway in cancer. Nat Rev Drug Discov 5:1026–1033
Sanchez P, Clement V, Altaba ARI (2005) Therapeutic targeting of the Hedgehog-GLI pathway in prostate cancer. Cancer Res 65:2990–2992
Smietana M, Kool ET (2002) Efficient and simple solid-phase synthesis of short cyclic oligodeoxynucleotides bearing a phosphorothioate linkage. Angew Chem Int Ed 41:3704–3707
van Beest M, Dooijes D, van de Wetering M, Kjaerulff S, Bonvin A, Nielsen O, Clevers H (2000) Sequence-specific high mobility group box factors recognize 10–12-base pair minor groove motifs. J Biol Chem 275:27266–27273
Van De Wetering M, Oosterwegel M, Dooijes D, Clevers H (1991) Identification and cloning of Tcf-1; a T lymphocyte-specific transcription factor containing a sequence-specific Hmg box. EMBO J 10:123–132
Acknowledgments
This research was funded by European Commission’s Sixth Framework Programme (Project reference AMNA, contract no. 013575). Oligonucleotides were synthesized by ATDBio Ltd. We are grateful to Professor Stefan Krauss, Jo Waaler and Martin Strand for carrying out the cell uptake studies.
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El-Sagheer, A.H., Brown, T. Synthesis, Serum Stability and Cell Uptake of Cyclic and Hairpin Decoy Oligonucleotides for TCF/LEF and GLI Transcription Factors. Int J Pept Res Ther 14, 367–372 (2008). https://doi.org/10.1007/s10989-008-9153-0
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DOI: https://doi.org/10.1007/s10989-008-9153-0