Generation of packaging cell lines for pseudotyped retroviral vectors of the G protein of vesicular stomatitis virus by using a modified tetracycline inducible system
Generation of packaging cell lines for pseudotyped retroviral vectors of the G protein of vesicular stomatitis virus by using a modified tetracycline inducible system
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DOI：
10.1073/pnas.93.19.10057
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发表时间：
1996-09-17
影响因子：
11.1
通讯作者：
Yee, JK
中科院分区：
文献类型：
--
作者：
Chen, ST;Iida, A;Yee, JK
我们已经证明水疱性口炎病毒(VSV-G)的G蛋白可以被整合到逆转录病毒的病毒粒子中，由于VSV-G的表达对大多数哺乳动物细胞是有毒的，建立稳定的VSV-G包装细胞系需要VSV-G的诱导表达，我们通过将雌激素受体的配体结合域融合到四环素调节的反式激活剂的羧基末端来修改四环素诱导系统。利用这个系统，我们发现VSV-G的表达是四环素依赖的，并且可以被β-雌二醇调节。稳定的包装细胞系可以很容易地建立起来，诱导VSV-G表达后可以产生酸性高滴度的伪型逆转录病毒载体。
We have previously shown that the G protein of vesicular stomatitis virus (VSV-G) can be incorporated into the virions of retroviruses, Since expression of VSV-G is toxic to most mammalian cells, development of stable VSV-G packaging cell lines requires inducible VSV-G expression, We have modified the tetracycline-inducible system by fusing the ligand binding domain of the estrogen receptor to the carboxy terminus of a tetracycline-regulated transactivator. Using this system, we show that VSV-G expression is tetracycline-dependent and can be modulated by beta-estradiol. Stable packaging cell lines can readily be established acid high-titer pseudotyped retroviral vectors can be generated upon induction of VSV-G expression.