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In this study, a method for assembling a nucleus in giant unilamellar vesicles (GUVs) is proposed. To induce reconstitution of the nucleus, the interphase egg extract of African clawed frogs\n                    <jats:italic>Xenopus laevis<\/jats:italic>\n                    is utilized, known as a biochemically controllable cell\u2010free system capable of transforming an added sperm chromatin into a nucleus in vitro. The GUV formation efficiency is enhanced by the inverted emulsion method through incorporating prolonged waiting time and adding chloroform into lipid\u2010dispersed oil, facilitating subsequent nuclear assembly reactions in the GUVs. Characterization of nucleus\u2010like structures formed in the GUVs revealed the presence of dense DNA and accumulated GFP\u2010NLS in the structure, indicative of functional nuclear import. Immunostaining further validated the presence of nuclear pore complexes on the surfaces of these nucleus\u2010like structures. The approach offers a versatile platform for constructing artificial cellular systems that closely mimic eukaryotic cells.\n                  <\/jats:p>","DOI":"10.1002\/smll.202412126","type":"journal-article","created":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T06:59:09Z","timestamp":1747724349000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Nuclear Assembly in Giant Unilamellar Vesicles Encapsulating\n                    <i>Xenopus<\/i>\n                    Egg Extract"],"prefix":"10.1002","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8892-2899","authenticated-orcid":false,"given":"Sho","family":"Takamori","sequence":"first","affiliation":[{"name":"Artificial Cell Membrane Systems Group Kanagawa Institute of Industrial Science and Technology  3\u20102\u20101 Sakado, Takatsu\u2010ku Kawasaki Kanagawa 213\u20100012 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