A SIMPLE METHOD FOR DELIVERING MORPHOLINO ANTISENSE OLIGOS INTO THE CYTOPLASM OF CELLS

We report a simple and effective means for delivering Morpholino antis ense oligos into the cytosol of cultured anchorage-dependent animal ce lls, This method, referred to as scrape-loading, is carried out in a m atter of seconds, uses a common inexpensive laboratory implement, and has minimal detrimental impact on the cells, Using this delivery metho d, a Morpholino oligo present at 0.1 mu M and 1 mu M in the extracellu lar medium inhibited its targeted genetic sequence within cultured Hel a 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 antisen se activity in cultured cells.