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タイトル: Saturation of transgene protein synthesis from mRNA in cells producing a large number of transgene mRNA.
著者: Takahashi, Yuki  kyouindb  KAKEN_id
Nishikawa, Makiya
Takiguchi, Naomi
Suehara, Tetsuya
Takakura, Yoshinobu  kyouindb  KAKEN_id
著者名の別形: 高橋, 有己
西川, 元也
キーワード: saturation
transgene expression
plasmid vector
mRNA expression
RNA interference
発行日: 2-May-2011
出版者: wiley
誌名: Biotechnology and bioengineering
巻: 108
号: 10
開始ページ: 2380
終了ページ: 2389
抄録: Experimental results have suggested that transgene expression can be saturated when large amounts of plasmid vectors are delivered into cells. To investigate this saturation kinetic behavior, cells were transfected with monitoring and competing plasmids using cationic liposomes. Even although an identical amount of a monitoring plasmid expressing firefly luciferase (FL) was used for transfection, transgene expression from the plasmid was greatly affected by the level of transgene expression from competing plasmids expressing renilla luciferase (RL). Similar results were obtained by exchanging the monitoring and competing plasmids. The competing plasmid-dependent reduction in transgene expression from the monitoring plasmid was also observed in mouse liver after hydrodynamic injection of plasmids. On the other hand, the mRNA and protein expression level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), an endogenous gene, in the liver hardly changed even when transgene expression process is saturated. The expression of FL from a monitoring plasmid was significantly restored by siRNA-mediated degradation of RL mRNA that was expressed from a competing plasmid. These results suggest that the efficiency of protein synthesis from plasmid vectors is reduced when a large amount of mRNA is transcribed with no significant changes in endogenous gene expression.
著作権等: This is the peer reviewed version of the following article: Takahashi, Y., Nishikawa, M., Takiguchi, N., Suehara, T. and Takakura, Y. (2011), Saturation of transgene protein synthesis from mRNA in cells producing a large number of transgene mRNA. Biotechnol. Bioeng., 108: 2380–2389, which has been published in final form at http://dx.doi.org/10.1002/bit.23179
This is not the published version. Please cite only the published version. 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。
URI: http://hdl.handle.net/2433/197140
DOI(出版社版): 10.1002/bit.23179
PubMed ID: 21520018
出現コレクション:学術雑誌掲載論文等

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