Introduction of antisense oligonucleotides into cells by permeabilization with streptolysin O.
Cellular uptake of antisense oligonucleotides is critical to their ability to inhibit gene expression. In the present study, phosphodiester oligodeoxynucleotides were introduced into cells during brief permeabilization with the pore-forming agent streptolysin O. The extent of antisense inhibition was dependent on the concentration of oligonucleotide present during permeabilization. In addition, the level of antisense inhibition was time-dependent; it reached a maximum at 18 h and subsequently diminished to control levels over the next 48 h. These results demonstrate the effectiveness of streptolysin O permeabilization as a means for simple and rapid introduction of oligonucleotides into eukaryotic cells.
Barry
E L
EL
Dartmouth Medical School, Hanover, NH.
Gesek
F A
FA
Friedman
P A
PA
D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels / Q000235:genetics*; D002461:Cell Line, Transformed; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002740:Chlorothiazide / Q000494:pharmacology; D007686:Kidney Tubules, Distal / Q000187:drug effects / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000378:metabolism*; D013301:Streptolysins / Q000494:pharmacology*
D001426:Bacterial Proteins; D015220:Calcium Channels; D016376:Oligonucleotides, Antisense; D013301:Streptolysins; C025964:streptolysin O; D002740:Chlorothiazide; D002118:Calcium