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Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved

Abstract

Efficient gene transfer and regulated transgene expression in primate embryonic stem (ES) cells are highly desirable for future applications of the cells. In the present study, we have examined using the nonintegrating Sendai virus (SeV) vector to introduce the green fluorescent protein (GFP) gene into non-human primate cynomolgus ES cells. The GFP gene was vigorously and stably expressed in the cynomolgus ES cells for a year. The cells were able to form fluorescent teratomas when transplanted into immunodeficient mice. They were also able to differentiate into fluorescent embryoid bodies, neurons, and mature blood cells. In addition, the GFP expression levels were reduced dose-dependently by the addition of an anti-RNA virus drug, ribavirin, to the culture. Thus, SeV vector will be a useful tool for efficient gene transfer into primate ES cells and the method of using antiviral drugs should allow further investigation for regulated SeV-mediated gene expression.

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Acknowledgements

Cynomolgus ES cells were provided by Norio Nakatsuji (Kyoto University, Kyoto, Japan), Yasushi Kondo (Tanabe Seiyaku Co. Ltd, Osaka, Japan), and Ryuzo Torii (Shiga University of Medical Science, Shiga, Japan). OP9 cells were provided by Toru Nakano (Osaka University, Osaka, Japan). We thank Yujiro Tanaka and Takayuki Asano for cultivating cynomolgus ES cells and Takeshi Hara for conducting NBT tests. We also thank Natsuko Kurosawa for technical assistance.

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Sasaki, K., Inoue, M., Shibata, H. et al. Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved. Gene Ther 12, 203–210 (2005). https://doi.org/10.1038/sj.gt.3302409

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