Plasmid DNA Entry Into Postmitotic Nuclei of Primary Rat Myotubes

• Published in: Proceedings of the National Academy of Sciences - PNAS 1995-05, Vol.92 (10), p.4572-4576
• Main Authors: Dowty, Martin E., Williams, Phillip, Zhang, Guofeng, Hagstrom, James E., Wolff, Jon A.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Base Sequence; beta-Galactosidase - biosynthesis; Cell Cycle; Cell nucleus; Cell Nucleus - metabolism; Cell Nucleus - ultrastructure; Cells, Cultured; Coleoptera - enzymology; Cytoplasm; Deoxyribonucleic acid; DNA; DNA Primers; Escherichia coli; Escherichia coli - enzymology; Gene therapy; Genetic Vectors; Genetics; Kinetics; Luciferases - biosynthesis; Microinjections; Mitosis; Molecular Sequence Data; Muscle fibers; Muscles - cytology; Muscles - metabolism; Nuclear pore; Plasmids; Plasmids - administration & dosage; Plasmids - metabolism; Proteins; Rats; Rats, Sprague-Dawley; RNA; Rodents; Sodium
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.92.10.4572

These studies were initiated to elucidate the mechanism of DNA nuclear transport in mammalian cells. Biotin- or gold-labeled plasmid and plasmid DNA expression vectors for Escherichia coli β-galactosidase or firefly luciferase were microinjected into the cytoplasm of primary rat myotubes in culture. Plasmid DNA was expressed in up to 70% of the injected myotubes, which indicates that it entered intact, postmitotic nuclei. The nuclear transport of plasmid DNA occurred through the nuclear pore by a process common to other large karyophilic macromolecules. The majority of the injected plasmid DNA was sequestered by cytoplasmic elements. This understanding of plasmid DNA nuclear transport provides a basis for increasing the efficiency of gene transfer.