The effects of N-terminal insertion into VSV-G of an scFv peptide.
The effects of N-terminal insertion into VSV-G of an scFv peptide.
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N末端插入SCFV肽的VSV-G的影响。
DOI：
10.1186/1743-422x-3-69
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发表时间：
2006-09-02
期刊：
影响因子：
4.8
通讯作者：
Piechaczyk, Marc
中科院分区：
文献类型：
--
作者：
Dreja, Hanna;Piechaczyk, Marc
重组逆转录病毒，包括慢病毒，是体外和体内稳定基因转移最广泛使用的载体。然而，无法选择性地将转基因传递到感兴趣的细胞中限制了该技术的使用。由于其广泛的趋向性、稳定性和假型一系列病毒载体的能力，水疱性口炎病毒 G 蛋白 (VSV-G) 是最常用的假型蛋白。在这里，我们尝试设计这种蛋白质以用于靶向目的。嵌合 VSV-G 蛋白是通过将细胞定向单链抗体 (scFv) 连接到其 N 末端而构建的。我们证明嵌合 VSV-G 分子可以整合到逆转录病毒和慢病毒颗粒中。用 VSV-G 假型化的 HIV-1 颗粒与针对人类主要组织相容性复合物 I 类 (MHC-I) 的 scFv 连接，与人类细胞强烈且特异性地结合。此外，这种新型分子优先驱动人类细胞的慢病毒转导，尽管滴度远低于用 VSV-G 假型化的病毒。这可能是由于修饰蛋白的融合活性低下。据我们所知，这是第一份成功设计VSV-G以包含大的（253个氨基酸）外源肽并尝试改变VSV-G假型载体的感染特征的报告。
Recombinant retroviruses, including lentiviruses, are the most widely used vectors for both in vitro and in vivo stable gene transfer. However, the inability to selectively deliver transgenes into cells of interest limits the use of this technology. Due to its wide tropism, stability and ability to pseudotype a range of viral vectors, vesicular stomatitis virus G protein (VSV-G) is the most commonly used pseudotyping protein. Here, we attempted to engineer this protein for targeting purposes. Chimaeric VSV-G proteins were constructed by linking a cell-directing single-chain antibody (scFv) to its N-terminal. We show that the chimaeric VSV-G molecules can integrate into retroviral and lentiviral particles. HIV-1 particles pseudotyped with VSV-G linked to an scFv against human Major Histocompatibility Complex class I (MHC-I) bind strongly and specifically to human cells. Also, this novel molecule preferentially drives lentiviral transduction of human cells, although the titre is considerably lower that viruses pseudotyped with VSV-G. This is likely due to the inefficient fusion activity of the modified protein. To our knowledge, this is the first report where VSV-G was successfully engineered to include a large (253 amino acids) exogenous peptide and where attempts were made to change the infection profile of VSV-G pseudotyped vectors.