A simple method for delivering morpholino antisense oligos into the cytoplasm of cells

Antisense Nucleic Acid Drug Dev. Fall 1996;6(3):169-75. doi: 10.1089/oli.1.1996.6.169.

Abstract

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.

MeSH terms

  • Animals
  • Base Sequence
  • Cell Line
  • Cytological Techniques*
  • Cytoplasm*
  • Enzyme Induction
  • HeLa Cells
  • Humans
  • Luciferases / biosynthesis
  • Molecular Sequence Data
  • Morpholines / chemistry*
  • Oligonucleotides, Antisense / administration & dosage*
  • Oligonucleotides, Antisense / chemistry
  • Plasmids / genetics
  • Rats
  • Sensitivity and Specificity
  • Time Factors
  • Transfection
  • Trypsin

Substances

  • Morpholines
  • Oligonucleotides, Antisense
  • Luciferases
  • Trypsin