title: Efficient gene delivery and expression in mammalian cells using DNA coupled with perfringolysin O.
abstract: Current non-viral DNA vectors for gene therapy are limited by low cellular transfection efficiencies and low levels of gene expression due to inefficient endosomal DNA release. We have used perfringolysin O (PFO), a membrane active bacterial protein, to deliver DNA into cells. PFO belongs to the so-called sulphydryl-activated family of membrane active bacterial proteins, which have been used to deliver small molecules and proteins into cells. PFO was incorporated into DNA complexes through a biotin-streptavidin bridge and the DNA-PFO complexes were used to deliver the Escherichia coli beta-galactosidase gene into cells. High levels of gene expression were achieved in murine sol 8 myoblast cells using these DNA-PFO complexes. The level of gene expression correlated well with the content of PFO in the complexes. Under optimal conditions, 15-20% of the cells were stained blue with X-gal. Furthermore, the expression was independent of a receptor ligand. Thus, membrane active bacterial proteins may be an important tool for the future development of non-viral DNA delivery systems for gene therapy.
authors.affiliation: Department of Cell Biology, Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: S
authors.initials: S
authors.lastname: Gottschalk
authors.forename: R K
authors.initials: RK
authors.lastname: Tweten
authors.forename: L C
authors.initials: LC
authors.lastname: Smith
authors.forename: S L
authors.initials: SL
authors.lastname: Woo
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000235:genetics / Q000494:pharmacology*; D002460:Cell Line; D004247:DNA / Q000235:genetics / Q000494:pharmacology*; D016503:Drug Delivery Systems; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006460:Hemolysin Proteins / Q000235:genetics / Q000494:pharmacology*; D051379:Mice; D001616:beta-Galactosidase / Q000235:genetics* / Q000494:pharmacology
chemical_list: D001427:Bacterial Toxins; D006460:Hemolysin Proteins; C020452:Clostridium perfringens theta-toxin; D004247:DNA; D001616:beta-Galactosidase