Plasmid DNA entry into postmitotic nuclei of primary rat myotubes.

作者：Martin E. Dowty, Phillip A. Williams, G Zhang, James E. Hagstrom, Jon A. Wolff · 发表于：Proceedings of the National Academy of Sciences · 年份：1995 · DOI：10.1073/pnas.92.10.4572 · 被引用次数：270 · 研究领域：Animal Genetics and Reproduction、CRISPR and Genetic Engineering、Virus-based gene therapy research

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.