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First published online March 29, 2007
Stem Cells Vol. 25 No. 7 July 2007, pp. 1707 -1712
doi:10.1634/stemcells.2006-0813; www.StemCells.com
© 2007 AlphaMed Press

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TECHNOLOGY DEVELOPMENT

Efficient Multicistronic Expression of a Transgene in Human Embryonic Stem Cells

Kouichi Hasegawaa,b, Aaron B. Cowanb, Norio Nakatsujib, Hirofumi Suemoria

aLaboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan;
bDepartment of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan

Key Words. Human embryonic stem cells • Genetic modification • Multicistronic expression

Correspondence: Hirofumi Suemori, D.Sc., Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawaharacho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Telephone: +81-75-751-3821; Fax: +81-75-751-3890; e-mail: hsuemori{at}frontier.kyoto-u.ac.jp

Received December 18, 2006; accepted for publication March 22, 2007.
First published online in STEM CELLS EXPRESS   March 29, 2007.



The applicability of human embryonic stem cells (hESCs) will be greatly enhanced by techniques that permit efficient genetic modification with multiple transgenes. We report here on single-promoter-driven foot-and-mouth disease virus segment 2A-mediated multicistronic expression of a transgene in hESCs. Efficient multicistronic expression of the transgene was permitted by 2A-mediated separation with almost the same amounts of encoded proteins in hESC. In addition, the multicistronic protein expression was successful in hESC-derived differentiated cells in in vivo and in vitro differentiation assays. This technology may be a significant advance in the genetic engineering of hESCs and hESC-derived cells for purposes that require the reliable expression of multiple transgenes.

Disclosure of potential conflicts of interest is found at the end of this article.






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