A simple method for delivering morpholino antisense oligos into the cytoplasm of cells.
We report a simple and effective means for delivering Morpholino antisense oligos into the cytosol of cultured anchorage-dependent animal cells. This method, referred to as scrape-loading, is carried out in a matter of seconds, uses a common inexpensive laboratory implement, and has minimal detrimental impact on the cells. Using this delivery method, a Morpholino oligo present at 0.1 microM and 1 microM in the extracellular medium inhibited its targeted genetic sequence within cultured Hela cells at levels of 56% and 85%, respectively. Lack of inhibition by two control Morpholino oligos at concentrations up to 3 microM indicates good sequence specificity by this structural type. Also described is a test system for simple, rapid, and sensitive quantitation of antisense activity in cultured cells.
ANTIVIRALS Inc., Corvallis, OR 97333, USA.
M
M
Partridge
A
A
Vincent
P
P
Matthews
J
J
Puma
D
D
Stein
J
J
Summerton
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003584:Cytological Techniques; D003593:Cytoplasm; D004790:Enzyme Induction; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D008969:Molecular Sequence Data; D009025:Morpholines / Q000737:chemistry*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000737:chemistry; D010957:Plasmids / Q000235:genetics; D051381:Rats; D012680:Sensitivity and Specificity; D013997:Time Factors; D014162:Transfection; D014357:Trypsin
D009025:Morpholines; D016376:Oligonucleotides, Antisense; D008156:Luciferases; D014357:Trypsin