Integration-free iPS cells engineered using human artificial chromosome vectors.
Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vectors, including episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3187830?pdf=render |
| _version_ | 1818527363376349184 |
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| author | Masaharu Hiratsuka Narumi Uno Kana Ueda Hajime Kurosaki Natsuko Imaoka Kanako Kazuki Etsuya Ueno Yutaro Akakura Motonobu Katoh Mitsuhiko Osaki Yasuhiro Kazuki Masato Nakagawa Shinya Yamanaka Mitsuo Oshimura |
| author_facet | Masaharu Hiratsuka Narumi Uno Kana Ueda Hajime Kurosaki Natsuko Imaoka Kanako Kazuki Etsuya Ueno Yutaro Akakura Motonobu Katoh Mitsuhiko Osaki Yasuhiro Kazuki Masato Nakagawa Shinya Yamanaka Mitsuo Oshimura |
| author_sort | Masaharu Hiratsuka |
| collection | DOAJ |
| description | Human artificial chromosomes (HACs) have unique characteristics as gene-delivery vectors, including episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown cassette. iHAC1 partially reprogrammed MEFs, and iHAC2 efficiently reprogrammed MEFs. Global gene expression patterns showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. Under non-selecting conditions, we established iHAC-free iPS cells by isolating cells that spontaneously lost iHAC2. Analyses of pluripotent markers, teratomas and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex virus thymidine kinase were eliminated by ganciclovir treatment, indicating that the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a built-in safeguard system. |
| first_indexed | 2024-12-11T06:35:08Z |
| format | Article |
| id | doaj.art-a312f18be0f84ff99bd61dcab579caa4 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-11T06:35:08Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-a312f18be0f84ff99bd61dcab579caa42022-12-22T01:17:23ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01610e2596110.1371/journal.pone.0025961Integration-free iPS cells engineered using human artificial chromosome vectors.Masaharu HiratsukaNarumi UnoKana UedaHajime KurosakiNatsuko ImaokaKanako KazukiEtsuya UenoYutaro AkakuraMotonobu KatohMitsuhiko OsakiYasuhiro KazukiMasato NakagawaShinya YamanakaMitsuo OshimuraHuman artificial chromosomes (HACs) have unique characteristics as gene-delivery vectors, including episomal transmission and transfer of multiple, large transgenes. Here, we demonstrate the advantages of HAC vectors for reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem (iPS) cells. Two HAC vectors (iHAC1 and iHAC2) were constructed. Both carried four reprogramming factors, and iHAC2 also encoded a p53-knockdown cassette. iHAC1 partially reprogrammed MEFs, and iHAC2 efficiently reprogrammed MEFs. Global gene expression patterns showed that the iHACs, unlike other vectors, generated relatively uniform iPS cells. Under non-selecting conditions, we established iHAC-free iPS cells by isolating cells that spontaneously lost iHAC2. Analyses of pluripotent markers, teratomas and chimeras confirmed that these iHAC-free iPS cells were pluripotent. Moreover, iHAC-free iPS cells with a re-introduced HAC encoding Herpes Simplex virus thymidine kinase were eliminated by ganciclovir treatment, indicating that the HAC safeguard system functioned in iPS cells. Thus, the HAC vector could generate uniform, integration-free iPS cells with a built-in safeguard system.http://europepmc.org/articles/PMC3187830?pdf=render |
| spellingShingle | Masaharu Hiratsuka Narumi Uno Kana Ueda Hajime Kurosaki Natsuko Imaoka Kanako Kazuki Etsuya Ueno Yutaro Akakura Motonobu Katoh Mitsuhiko Osaki Yasuhiro Kazuki Masato Nakagawa Shinya Yamanaka Mitsuo Oshimura Integration-free iPS cells engineered using human artificial chromosome vectors. PLoS ONE |
| title | Integration-free iPS cells engineered using human artificial chromosome vectors. |
| title_full | Integration-free iPS cells engineered using human artificial chromosome vectors. |
| title_fullStr | Integration-free iPS cells engineered using human artificial chromosome vectors. |
| title_full_unstemmed | Integration-free iPS cells engineered using human artificial chromosome vectors. |
| title_short | Integration-free iPS cells engineered using human artificial chromosome vectors. |
| title_sort | integration free ips cells engineered using human artificial chromosome vectors |
| url | http://europepmc.org/articles/PMC3187830?pdf=render |
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