Biopolym. Cell. 2008; 24(2):171-175.
http://dx.doi.org/10.7124/bc.00079E
Bioorganic Chemistry
Synthesis of fluorescently labeled oligonucleotide conjugate with transport peptide on modified Silochrom-2 support
1Dubey I. Ya.
  1. Institute of Molecular Biology and Genetics, NAS of Ukraine
    150, Akademika Zabolotnoho Str., Kyiv, Ukraine, 03680

Abstract

Synthesis of triple conjugate of oligonucleotide with transport peptide and fluorophore is described. 5'-aminoalkyl-modified 3'-fluorescein-labeled 18-mer oligonucleotide, complementary to the region of E. coli β-galactosidase gene was prepared by solid phase synthesis on fluorescein-modified support Silochrom-2. Its treatment with iodoacetic anhydride led to the formation of iodoacetamide derivative. The latter reacted specifically with cystein-containing 10-mer peptide, a fragment of the transport domain of large T-antigen of SV40 virus, to provide high yield of target hybrid.
Keywords: oligonucleotide conjugates, transport peptides, fluorescein
Full text: (PDF, in Ukrainian)

References

[1] Fischer PM, Krausz E, Lane DP. Cellular delivery of impermeable effector molecules in the form of conjugates with peptides capable of mediating membrane translocation. Bioconjug Chem. 2001;12(6):825-41.
[2] Pierce TL, White AR, Tregear GW, Sexton PM. Peptide-oligonucleotide hybrids in antisense therapy. Mini Rev Med Chem. 2005;5(1):41-55.
[3] Dubey I, Pratviel G, Meunier B. Modification of the thiourea linkage of a fluorescein-oligonucleotide conjugate to a guanidinium motif during ammonia deprotection. Bioconjug Chem. 1998;9(5):627-32.
[4] Dubey I. Ya., Dubey L. V., Fedoryak D. M. Synthesis of 3'- and 3',5'-modified oligonucleotides on functionalized silica Silochrom-2. Biopolym. Cell. 2007; 23(2):137-142
[5] Bongartz JP, Aubertin AM, Milhaud PG, Lebleu B. Improved biological activity of antisense oligonucleotides conjugated to a fusogenic peptide. Nucleic Acids Res. 1994;22(22):4681-8.
[6] Turner JJ, Arzumanov AA, Gait MJ. Synthesis, cellular uptake and HIV-1 Tat-dependent trans-activation inhibition activity of oligonucleotide analogues disulphide-conjugated to cell-penetrating peptides. Nucleic Acids Res. 2005;33(1):27-42.
[7] Reed MW, Fraga D, Schwartz DE, Scholler J, Hinrichsen RD. Synthesis and evaluation of nuclear targeting peptide-antisense oligodeoxynucleotide conjugates. Bioconjug Chem. 1995;6(1):101-8.
[8] Eritja R., Pons A., Escarceller M., Giralt E., Albericio F. Synthesis of defined peptide-oligonucleotide hybrids containing a nuclear transport signal sequence. Tetrahedron. 1991; 47(24):4113–4120.
[9] de La Torre BG, Albericio F, Saison-Behmoaras E, Bachi A, Eritja R. Synthesis and binding properties of oligonucleotides carrying nuclear localization sequences. Bioconjug Chem. 1999;10(6):1005-12.
[10] Handbook of biochemistry and molecular biology. Ed. G. Fasman. Boca Raton: CRC press, 1975. Vol. 1:175.