Hairpin, dumbbell, and single-stranded phosphodiester oligonucleotides exhibit identical uptake in T lymphocyte cell lines.
The cellular binding, uptake, and intracellular distribution of structured double-stranded phosphodiester oligonucleotides (decoys) have been examined in T lymphocytes using fluorescein-labeled molecules. Intracellular localization of hairpin and dumbbell decoys was similar to that of single-stranded oligonucleotides. At short incubation times, oligonucleotides were localized only in cytoplasmic vesicles, whereas at longer times, they were also found in the nucleus. Cellular uptake was dependent on temperature, time, and extracellular concentration. Oligonucleotide efflux was similar for all types of molecules and was very rapid (t1/2 = 10-15 minutes). These results imply that phosphodiester double-stranded oligonucleotides can enter cellular compartments of interest for their potential biologic function and, thus, provide a potential tool to control gene transcription.
GENSET, Paris, France.
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L
Aguilar
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A
Hémar
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A
Dautry-Varsat
M
M
Blumenfeld
D004277:DNA, Single-Stranded / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics; D004952:Esters; D005089:Exocytosis; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D006801:Humans; D019169:Jurkat Cells; D018613:Microscopy, Confocal; D009690:Nucleic Acid Conformation; D009841:Oligonucleotides / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics; D063065:Organophosphonates / Q000097:blood / Q000737:chemistry* / Q000493:pharmacokinetics; D013601:T-Lymphocytes / Q000378:metabolism*
D004277:DNA, Single-Stranded; D004952:Esters; D009841:Oligonucleotides; D063065:Organophosphonates; D016650:Fluorescein-5-isothiocyanate