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Phosphorothioate Antisense Oligonucleotides Induce the Formation of Nuclear Bodies

Authors :
Lorenz, Peter
Baker, Brenda F.
Bennett, C. Frank
Spector, David L.
Source :
Molecular Biology of the Cell; May 1998, Vol. 9 Issue: 5 p1007-1023, 17p
Publication Year :
1998

Abstract

Antisense oligonucleotides are powerful tools for the in vivo regulation of gene expression. We have characterized the intracellular distribution of fluorescently tagged phosphorothioate oligodeoxynucleotides (PS-ONs) at high resolution under conditions in which PS-ONs have the potential to display antisense activity. Under these conditions PS-ONs predominantly localized to the cell nucleus where they accumulated in 20–30 bright spherical foci designated phosphorothioate bodies (PS bodies), which were set against a diffuse nucleoplasmic population excluding nucleoli. PS bodies are nuclear structures that formed in cells after PS-ON delivery by transfection agents or microinjection but were observed irrespectively of antisense activity or sequence. Ultrastructurally, PS bodies corresponded to electron-dense structures of 150–300 nm diameter and resembled nuclear bodies that were found with lower frequency in cells lacking PS-ONs. The environment of a living cell was required for the de novo formation of PS bodies, which occurred within minutes after the introduction of PS-ONs. PS bodies were stable entities that underwent noticeable reorganization only during mitosis. Upon exit from mitosis, PS bodies were assembled de novo from diffuse PS-ON pools in the daughter nuclei. In situ fractionation demonstrated an association of PS-ONs with the nuclear matrix. Taken together, our data provide evidence for the formation of a nuclear body in cells after introduction of phosphorothioate oligodeoxynucleotides.

Details

Language :
English
ISSN :
10591524 and 19394586
Volume :
9
Issue :
5
Database :
Supplemental Index
Journal :
Molecular Biology of the Cell
Publication Type :
Periodical
Accession number :
ejs46606692
Full Text :
https://doi.org/10.1091/mbc.9.5.1007