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j.1365-2443.2011.01528.x.pdf | 1.02 MB | Adobe PDF | 見る/開く |
タイトル: | Simple and efficient method for generation of induced pluripotent stem cells using piggyBac transposition of doxycycline-inducible factors and an EOS reporter system. |
著者: | Tsukiyama, Tomoyuki Asano, Ryota Kawaguchi, Takamasa Kim, Narae Yamada, Masayasu Minami, Naojiro Ohinata, Yasuhide Imai, Hiroshi |
著者名の別形: | 今井, 裕 |
発行日: | 9-Jun-2011 |
出版者: | wiley |
誌名: | Genes to Cells |
巻: | 16 |
号: | 7 |
開始ページ: | 815 |
終了ページ: | 825 |
抄録: | PiggyBac (PB) transposition of reprogramming factors (Oct3/4 (O), Sox2 (S), Klf4 (K) and c-Myc) is a safe, nonviral method for generating induced pluripotent stem cells (iPSCs). However, compared with retroviral methods, the reprogramming efficiency of the PB-mediated methods is relatively low. In this study, we describe a simple and efficient system for generating high-quality iPSCs by a single transfection of multiple plasmids that does not require the use of a virus, special instruments or skilled techniques. To improve reprogramming efficiency, we modified the components of the polycistronic 2A vectors used in this study and also investigated the combination of another reprogramming-related factor (L-Myc). By simultaneous transposition of multiple PB vectors containing an EOS (early transposon promoter and Oct3/4 and Sox2 enhancers) reporter and modified polycistronic doxycycline (Dox)-inducible factors, we reprogrammed mouse somatic cells with an efficiency higher than is usually obtained with retroviral methods and we established some iPSC lines that contributed highly to chimeras. By using the Dox-inducible system, we also showed that the appropriate elimination of exogenous-factor expression at appropriate time accelerated the induction of Oct3/4 when a combination of OKS and c-Myc vectors were used. |
著作権等: | This is the peer reviewed version of the following article: Tsukiyama, T., Asano, R., Kawaguchi, T., Kim, N., Yamada, M., Minami, N., Ohinata, Y. and Imai, H. (2011), Simple and efficient method for generation of induced pluripotent stem cells using piggyBac transposition of doxycycline-inducible factors and an EOS reporter system. Genes to Cells, 16: 815–825, which has been published in final form at http://dx.doi.org/10.1111/j.1365-2443.2011.01528.x この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。 This is not the published version. Please cite only the published version. |
URI: | http://hdl.handle.net/2433/197277 |
DOI(出版社版): | 10.1111/j.1365-2443.2011.01528.x |
PubMed ID: | 21658168 |
出現コレクション: | 学術雑誌掲載論文等 |
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