Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
| Autor: | Tatsuo Fukagawa, Tokuko Haraguchi, Yasushi Hiraoka, Yuji Chikashige, Yasuhiro Hirano, Hiroko Osakada, Shouhei Kobayashi, Yasushi Okada, Tomoko Shindo, Shinsuke Shibata, Sukriye Bilir, Haruhiko Asakawa, Takako Koujin, Chie Mori, Kohei Nishimura |
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| Rok vydání: | 2021 |
| Předmět: |
animal structures
QH301-705.5 viruses Medicine (miscellaneous) Cellular imaging Transfection General Biochemistry Genetics and Molecular Biology Article Nuclear envelope Plasmid dna Cell Line Tumor medicine Humans Telophase Biology (General) health care economics and organizations Envelope (waves) Cell Nucleus Chemistry fungi Membrane Proteins Nuclear Proteins Biological Transport DNA Cell biology medicine.anatomical_structure Gene Expression Regulation embryonic structures Mutation Single-Cell Analysis General Agricultural and Biological Sciences Nucleus Plasmids |
| Zdroj: | Communications Biology Communications Biology, Vol 5, Iss 1, Pp 1-15 (2022) |
| ISSN: | 2399-3642 |
| Popis: | DNA transfection is an important technology in life sciences, wherein nuclear entry of DNA is necessary to express exogenous DNA. Non-viral vectors and their transfection reagents are useful as safe transfection tools. However, they have no effect on the transfection of non-proliferating cells, the reason for which is not well understood. This study elucidates the mechanism through which transfected DNA enters the nucleus for gene expression. To monitor the behavior of transfected DNA, we introduce plasmid bearing lacO repeats and RFP-coding sequences into cells expressing GFP-LacI and observe plasmid behavior and RFP expression in living cells. RFP expression appears only after mitosis. Electron microscopy reveals that plasmids are wrapped with nuclear envelope (NE)‒like membranes or associated with chromosomes at telophase. The depletion of BAF, which is involved in NE reformation, delays plasmid RFP expression. These results suggest that transfected DNA is incorporated into the nucleus during NE reformation at telophase. Haraguchi et al. investigate how transfected DNA is incorporated into the nucleus using light and electron microscopy, and the LacI/LacO system. The authors report that cytoplasmic localised plasmid DNA is incorporated into the nucleus as it reforms during M-exit and transgene expression was achieved only after nuclear reformation in the next G1. |
| Databáze: | OpenAIRE |
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