Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase

Journal	Commun. Biol. 5(1):78 (2022)
Title	Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
Laboratory	Nuclear Dynamics Group

Abstract

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.

Authors	Tokuko Haraguchi (1, 2), Takako Koujin (1, 2), Tomoko Shindo (3), Şükriye Bilir (1), Hiroko Osakada (2), Kohei Nishimura (1), Yasuhiro Hirano (1), Haruhiko Asakawa (1), Chie Mori (2), Shouhei Kobayashi (2), Yasushi Okada (4, 5), Yuji Chikashige (2), Tatsuo Fukagawa (1), Shinsuke Shibata (3), Yasushi Hiraoka (1, 2) Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita 565-0871, Japan. Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan. Keio University, School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan. RIKEN Center for Biosystems Dynamics Research (BDR), Suita, Osaka 565-0874, Japan. Department of Cell Biology, Department of Physics, Universal Biology Institute (UBI) and International Research Center for Neurointelligence (IRCN), the University of Tokyo, Tokyo 113-0033, Japan.
PubMed	35058555

Authors

Tokuko Haraguchi (1, 2), Takako Koujin (1, 2), Tomoko Shindo (3), Şükriye Bilir (1), Hiroko Osakada (2), Kohei Nishimura (1), Yasuhiro Hirano (1), Haruhiko Asakawa (1), Chie Mori (2), Shouhei Kobayashi (2), Yasushi Okada (4, 5), Yuji Chikashige (2), Tatsuo Fukagawa (1), Shinsuke Shibata (3), Yasushi Hiraoka (1, 2)

Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka, Suita 565-0871, Japan.
Advanced ICT Research Institute Kobe, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan.
Keio University, School of Medicine, Shinjuku-ku, Tokyo 160-8582, Japan.
RIKEN Center for Biosystems Dynamics Research (BDR), Suita, Osaka 565-0874, Japan.
Department of Cell Biology, Department of Physics, Universal Biology Institute (UBI) and International Research Center for Neurointelligence (IRCN), the University of Tokyo, Tokyo 113-0033, Japan.

PubMed

35058555