1. Oligodeoxyribonucleotide Phosphorothioate Fluxes and Localization in Hematopoietic Cells
- Author
-
Makoto Matsukura, Samuel Broder, William Egan, Gerald E. Marti, Gerald Zon, and Philip D. Noguchi
- Subjects
Molecular Sequence Data ,Cell ,Biology ,Cell Line ,Flow cytometry ,Tumor Cells, Cultured ,Genetics ,medicine ,Fluorescence microscope ,Humans ,Base Sequence ,medicine.diagnostic_test ,Oligonucleotide ,Organothiophosphates ,Biological Transport ,Oligonucleotides, Antisense ,Flow Cytometry ,Molecular biology ,Kinetics ,Haematopoiesis ,Cell nucleus ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Indicators and Reagents ,Efflux ,Intracellular ,HeLa Cells - Abstract
An antisense oligonucleotide phosphorothioate, previously shown to inhibit HIV-1 viral expression in chronically infected H9 cells, was fluorescently labeled to study oligonucleotide fluxes and localization within living cells. Observations based on flow cytometry and fluorescence microscopy show the following: within around 0.5-2 h, an apparent steady-state distribution of the oligonucleotide is achieved in which the intracellular oligonucleotide concentration is less than that present in the external medium; following oligonucleotide uptake and resuspension of the cells in oligonucleotide-free medium, an oligonucleotide efflux, with a time constant similar to that for uptake, is observed (although a significant fraction of the phosphorothioate remains within the cell); cellular uptake as a function of the external oligonucleotide concentration is nonlinear, being more efficient at lower concentrations (less than 2 microM); and a predominant oligonucleotide localization within the cell nucleus and perinuclear organelles is observed.
- Published
- 1992