Dreja Hanna, Piechaczyk Marc
Institut de Génétique Moléculaire de Montpellier, UMR 5535, IFR122, CNRS, France.
Virol J. 2006 Sep 2;3:69. doi: 10.1186/1743-422X-3-69.
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.
重组逆转录病毒，包括慢病毒，是体外和体内稳定基因转移中使用最广泛的载体。然而，无法将转基因选择性地递送至感兴趣的细胞限制了这项技术的应用。由于其广泛的嗜性、稳定性以及能够对一系列病毒载体进行假型化，水泡性口炎病毒G蛋白（VSV-G）是最常用的假型化蛋白。在此，我们试图对该蛋白进行工程改造以用于靶向目的。通过将细胞导向性单链抗体（scFv）连接到其N端构建嵌合VSV-G蛋白。我们表明嵌合VSV-G分子可以整合到逆转录病毒和慢病毒颗粒中。用与针对人类主要组织相容性复合体I类（MHC-I）的scFv连接的VSV-G假型化的HIV-1颗粒能强烈且特异性地与人细胞结合。此外，这种新型分子优先驱动人细胞的慢病毒转导，尽管其滴度比用VSV-G假型化的病毒低得多。这可能是由于修饰蛋白的融合活性效率低下所致。据我们所知，这是首次成功将VSV-G工程改造以包含一个大的（253个氨基酸）外源肽并尝试改变VSV-G假型化载体感染谱的报告。
