Gene introduction into the mitochondria of Arabidopsis thaliana via peptide-based carriers

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 mitochond...

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Bibliographic Details
Published in:Scientific reports 2015-01, Vol.5 (1), p.7751-7751, Article 7751
Main Authors: Chuah, Jo-Ann, Yoshizumi, Takeshi, Kodama, Yutaka, Numata, Keiji
Format: Article
Language:English
Subjects:
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Amino Acid Sequence
Arabidopsis - genetics
Blotting, Western
Cell-Penetrating Peptides - metabolism
Deoxyribonucleic acid
DNA
DNA, Plant - metabolism
Electrostatic properties
Gene transfer
Gene Transfer Techniques
Genes, Plant
Genetic transformation
Genetic Vectors - metabolism
Green Fluorescent Proteins - metabolism
Humanities and Social Sciences
Internalization
Intracellular
Mitochondria
Mitochondria - metabolism
Mitochondrial DNA
Molecular Sequence Data
multidisciplinary
Peptides
Peptides - chemistry
Peptides - metabolism
Plant Leaves - metabolism
Plants, Genetically Modified
Plasmids - metabolism
Science
Transfection
Transformation, Genetic
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep07751