title: Mitochondria transfection by oligonucleotides containing a signal peptide and vectorized by cationic liposomes.
abstract: The progress of research in gene therapy allows hope for treatment of mitochondrial genetic disorders provided that efficient methods for gene transfer into mitochondria can be found. In this work, we have used an oligonucleotide coupled covalently to a mitochondria-targeted peptide at one end and a cationic liposome prepared from trimethyl aminoethane carbamoyl cholesterol iodide (TMAEC-Chol) to carry it in living cells. With a fluorescent probe to label the oligonucleotide at the other end and by means of confocal microscopy, we show that such modified oligonucleotides complexed to liposomes enter into the cytoplasm of human fibroblasts in primary culture, and then, after dissociation from the complexes, they penetrate into the mitochondria. The fluorescence was still observed after 8 days, suggesting the continued presence of oligonucleotides. At the concentrations used for this study, the cationic liposomes have practically no effect on cell growth, as revealed by the MTT assay.
authors.affiliation: INSERM U393, Unité de Reserche sur les Handicaps Génétiqies de l'Enfant, Hĵpital Necker-Enfants Malades, Paris, France.
authors.forename: V
authors.initials: V
authors.lastname: Geromel
authors.forename: A
authors.initials: A
authors.lastname: Cao
authors.forename: D
authors.initials: D
authors.lastname: Briane
authors.forename: J
authors.initials: J
authors.lastname: Vassy
authors.forename: A
authors.initials: A
authors.lastname: Rotig
authors.forename: P
authors.initials: P
authors.lastname: Rustin
authors.forename: R
authors.initials: R
authors.lastname: Coudert
authors.forename: J P
authors.initials: JP
authors.lastname: Rigaut
authors.forename: A
authors.initials: A
authors.lastname: Munnich
authors.forename: E
authors.initials: E
authors.lastname: Taillandier
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002412:Cations; D002478:Cells, Cultured; D019793:Fluorescein; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008081:Liposomes; D008928:Mitochondria / Q000235:genetics*; D028361:Mitochondrial Diseases / Q000235:genetics / Q000628:therapy; D008969:Molecular Sequence Data; D020319:Oligodeoxyribonucleotides, Antisense / Q000008:administration & dosage / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics*; D014162:Transfection
chemical_list: D002412:Cations; D008081:Liposomes; D020319:Oligodeoxyribonucleotides, Antisense; D021382:Protein Sorting Signals; D019793:Fluorescein