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 mu M and 1 mu M 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 mu M 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.