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

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...

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Autores principales: Haraguchi, Tokuko, Koujin, Takako, Shindo, Tomoko, Bilir, Şükriye, Osakada, Hiroko, Nishimura, Kohei, Hirano, Yasuhiro, Asakawa, Haruhiko, Mori, Chie, Kobayashi, Shouhei, Okada, Yasushi, Chikashige, Yuji, Fukagawa, Tatsuo, Shibata, Shinsuke, Hiraoka, Yasushi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8776997/
https://www.ncbi.nlm.nih.gov/pubmed/35058555
http://dx.doi.org/10.1038/s42003-022-03021-8
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author Haraguchi, Tokuko
Koujin, Takako
Shindo, Tomoko
Bilir, Şükriye
Osakada, Hiroko
Nishimura, Kohei
Hirano, Yasuhiro
Asakawa, Haruhiko
Mori, Chie
Kobayashi, Shouhei
Okada, Yasushi
Chikashige, Yuji
Fukagawa, Tatsuo
Shibata, Shinsuke
Hiraoka, Yasushi
author_facet Haraguchi, Tokuko
Koujin, Takako
Shindo, Tomoko
Bilir, Şükriye
Osakada, Hiroko
Nishimura, Kohei
Hirano, Yasuhiro
Asakawa, Haruhiko
Mori, Chie
Kobayashi, Shouhei
Okada, Yasushi
Chikashige, Yuji
Fukagawa, Tatsuo
Shibata, Shinsuke
Hiraoka, Yasushi
author_sort Haraguchi, Tokuko
collection PubMed
description 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.
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spelling pubmed-87769972022-02-04 Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase Haraguchi, Tokuko Koujin, Takako Shindo, Tomoko Bilir, Şükriye Osakada, Hiroko Nishimura, Kohei Hirano, Yasuhiro Asakawa, Haruhiko Mori, Chie Kobayashi, Shouhei Okada, Yasushi Chikashige, Yuji Fukagawa, Tatsuo Shibata, Shinsuke Hiraoka, Yasushi Commun Biol Article 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. Nature Publishing Group UK 2022-01-20 /pmc/articles/PMC8776997/ /pubmed/35058555 http://dx.doi.org/10.1038/s42003-022-03021-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Haraguchi, Tokuko
Koujin, Takako
Shindo, Tomoko
Bilir, Şükriye
Osakada, Hiroko
Nishimura, Kohei
Hirano, Yasuhiro
Asakawa, Haruhiko
Mori, Chie
Kobayashi, Shouhei
Okada, Yasushi
Chikashige, Yuji
Fukagawa, Tatsuo
Shibata, Shinsuke
Hiraoka, Yasushi
Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
title Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
title_full Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
title_fullStr Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
title_full_unstemmed Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
title_short Transfected plasmid DNA is incorporated into the nucleus via nuclear envelope reformation at telophase
title_sort transfected plasmid dna is incorporated into the nucleus via nuclear envelope reformation at telophase
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8776997/
https://www.ncbi.nlm.nih.gov/pubmed/35058555
http://dx.doi.org/10.1038/s42003-022-03021-8
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