Achieving efficient delivery of morpholino oligos in cultured cells.
One of the many features that make morpholino oligos unique among the antisense structural types is an uncharged backbone. While this feature eliminates the nonspecific interactions of traditional S-oligos, it also renders the morpholino undeliverable via the traditional lipid-based delivery systems. This article describes a highly efficient method of delivering morpholino oligos into adherent and nonadherent cultured cells. In this system, a nonionic morpholino oligo is paired to a complementary DNA "carrier." The DNA is then bound electrostatically to a partially ionized, weakly-basic ethoxylated polyethylenimine (EPEI). This morpholino/DNA/EPEI complex is efficiently endocytosed, and when the pH drops within the endosome, the EPEI more fully ionizes, resulting in permeabilization of the endosomal membrane and release of the morpholino into the cytosol. This article describes optimization of delivery in HeLa cells and provides the basis for delivery in any cultured endocytic cell type. genesis 30:94--102, 2001.
Morcos
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GENE TOOLS, LLC, Philomath, Oregon 97370, USA. pmorcos@gene-tools.com
D001483:Base Sequence; D002448:Cell Adhesion; D004247:DNA / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009025:Morpholines / Q000008:administration & dosage / Q000378:metabolism*; D009841:Oligonucleotides / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011094:Polyethyleneimine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D012680:Sensitivity and Specificity; D055672:Static Electricity; D013997:Time Factors; D014162:Transfection / Q000379:methods
D009025:Morpholines; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D011094:Polyethyleneimine; D004247:DNA