Triple, MPEG-conjugated, helix-forming oligonucleotides (TRIPEGXs): liquid-phase synthesis of natural and chimeric "all-purine" sequences linked to high molecular weight poly(ethylene glycols).
Long "all-purine" oligonucleotides, up to the 20mer, known to be active as antigene effectors, conjugated to high molecular weight monomethoxy poly(ethylene glycol)s (MPEG)s, were successfully synthesized. Through a liquid-phase, MPEG-supported process, both natural and chimeric sequences containing selected phosphorothioate backbone modifications were obtained, purified, and characterized. To follow their cellular trafficking, a fluorescent probe was linked by soluble supported organic reactions to the 5'-terminus, and the efficiency of the different synthetic procedures for the introduction of a fluorescein moiety was compared. The usefulness of the fluorescent marker was estimated by laser confocal microscopy that ascertains that the MPEG-conjugation enhances the oligonucleotide capacity to cross the cellular membranes and to be accumulated inside the nuclei.
Ballico
M
M
Department of Chemical Science, Via Giorgieri 1, University of Trieste, 34127 Trieste, Italy.
Drioli
S
S
Morvan
F
F
Xodo
L
L
Bonora
G M
GM
D002463:Cell Membrane Permeability; D004247:DNA; D004337:Drug Carriers / Q000737:chemistry / Q000493:pharmacokinetics; D019793:Fluorescein; D005456:Fluorescent Dyes; D006801:Humans; D020014:K562 Cells; D018613:Microscopy, Confocal; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000493:pharmacokinetics*; D010710:Phosphates; D011092:Polyethylene Glycols / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D011687:Purines / Q000737:chemistry; D013873:Thionucleotides
D004337:Drug Carriers; D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D010710:Phosphates; D011687:Purines; D013873:Thionucleotides; C051802:triplex DNA; D011092:Polyethylene Glycols; C083120:phosphorodithioic acid; C028210:monomethoxypolyethylene glycol; D004247:DNA; D019793:Fluorescein