Plasmid DNA entry into postmitotic nuclei of primary rat myotubes.
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 beta-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.
Dowty
M E
ME
Department of Pediatrics, Waisman Center, University of Wisconsin, Madison 53705, USA.
Williams
P
P
Zhang
G
G
Hagstrom
J E
JE
Wolff
J A
JA
D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D001517:Coleoptera / Q000201:enzymology; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology; D005822:Genetic Vectors; D007700:Kinetics; D008156:Luciferases / Q000096:biosynthesis; D008845:Microinjections; D008938:Mitosis; D008969:Molecular Sequence Data; D009132:Muscles / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D001616:beta-Galactosidase / Q000096:biosynthesis
D017931:DNA Primers; D008156:Luciferases; D001616:beta-Galactosidase