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Start Over Author "Kana Ueda" Publisher public library of science (plos)
2 results on '"Kana Ueda"'

1. Integration-free iPS cells engineered using human artificial chromosome vectors.

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, and Mitsuo Oshimura

Subjects
Medicine, Science
Abstract

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.

Published
2011
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2. Integration-free iPS cells engineered using human artificial chromosome vectors

Author

Yutaro Akakura, Mitsuhiko Osaki, Masato Nakagawa, Narumi Uno, Kana Ueda, Natsuko Imaoka, Kanako Kazuki, Masaharu Hiratsuka, Mitsuo Oshimura, Etsuya Ueno, Motonobu Katoh, Hajime Kurosaki, Shinya Yamanaka, and Yasuhiro Kazuki

Subjects
Transgene, Genetic Vectors, Induced Pluripotent Stem Cells, Biomedical Engineering, Gene Expression, lcsh:Medicine, Bioengineering, Human artificial chromosome, CHO Cells, Biology, Thymidine Kinase, Chromosomes, Artificial, Human, Mice, Cricetulus, Cricetinae, Gene expression, Molecular Cell Biology, Animals, Humans, Simplexvirus, Jian R. Lu, Vector (molecular biology), Induced pluripotent stem cell, lcsh:Science, Cell Engineering, Multidisciplinary, Stem Cells, lcsh:R, Genes, Transgenic, Suicide, Fibroblasts, Embryonic stem cell, Molecular biology, The University of Manchester, United Kingdom, Thymidine kinase, lcsh:Q, Cellular Types, Genetic Engineering, Reprogramming, Research Article, Biotechnology
Abstract

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.

Published
2011

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