title: Gene introduction into the mitochondria of Arabidopsis thaliana via peptide-based carriers.
abstract: Available methods in plant genetic transformation are nuclear and plastid transformations because similar procedures have not yet been established for the mitochondria. The double membrane and small size of the organelle, in addition to its large population in cells, are major obstacles in mitochondrial transfection. Here we report the intracellular delivery of exogenous DNA localized to the mitochondria of Arabidopsis thaliana using a combination of mitochondria-targeting peptide and cell-penetrating peptide. Low concentrations of peptides were sufficient to deliver DNA into the mitochondria and expression of imported DNA reached detectable levels within a short incubation period (12 h). We found that electrostatic interaction with the cell membrane is not a critical factor for complex internalization, instead, improved intracellular penetration of mitochondria-targeted complexes significantly enhanced gene transfer efficiency. Our results delineate a simple and effective peptide-based method, as a starting point for the development of more sophisticated plant mitochondrial transfection strategies.
authors.affiliation: Enzyme Research Team, Biomass Engineering Program Cooperation Division, Center for Sustainable Resource Science, RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
authors.forename: Jo-Ann
authors.initials: JA
authors.lastname: Chuah
authors.affiliation: Institute for Advanced Biosciences, Keio University, 403-1 Nipponkoku, Daihoji, Tsuruoka, Yamagata 997-0017, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yoshizumi
authors.affiliation: Center for Bioscience Research and Education, Utsunomiya University, Tochigi 321-8505, Japan.
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Kodama
authors.affiliation: Enzyme Research Team, Biomass Engineering Program Cooperation Division, Center for Sustainable Resource Science, RIKEN, 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan.
authors.forename: Keiji
authors.initials: K
authors.lastname: Numata
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D015153:Blotting, Western; D057846:Cell-Penetrating Peptides / Q000378:metabolism; D018744:DNA, Plant / Q000378:metabolism; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005822:Genetic Vectors / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D018515:Plant Leaves / Q000378:metabolism; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D057846:Cell-Penetrating Peptides; D018744:DNA, Plant; D010455:Peptides; D049452:Green Fluorescent Proteins