Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved
Efficient and stable Sendai virus-mediated gene transfer into primate embryonic stem cells with pluripotency preserved
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DOI：
10.1038/sj.gt.3302409
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发表时间：
2005-02-01
期刊：
影响因子：
5.1
通讯作者：
Hanazono, Y
中科院分区：
文献类型：
--
作者：
Sasaki, K;Inoue, M;Hanazono, Y
在灵长类动物胚胎干细胞中进行有效的基因转移和受调控的转基因表达对于该细胞的未来应用是高度期望的。在本研究中，我们已经检查了使用非整合仙台病毒（SeV）载体引入绿色荧光蛋白（GFP）基因到非人灵长类食蟹猴ES细胞。GFP基因在食蟹猴ES细胞中强烈且稳定地表达了一年。当移植到免疫缺陷小鼠体内时，这些细胞能够形成荧光畸胎瘤。它们还能够分化成荧光胚状体、神经元和成熟血细胞。此外，通过向培养物中加入抗RNA病毒药物利巴韦林，GFP表达水平呈剂量依赖性降低。因此，SeV载体将是一个有用的工具，有效的基因转移到灵长类动物ES细胞和使用抗病毒药物的方法，应允许进一步调查调节SeV介导的基因表达。
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.