Logvinoff C, Epstein A L
Centre de Génétique Moléculaire et Cellulaire, CNRS-UMR 5534, Université Claude Bernard Lyon 1, 69622 Villeurbanne Cedex, France.
Hum Gene Ther. 2001 Jan 20;12(2):161-7. doi: 10.1089/104303401750061221.
Helper-dependent HSV vectors (commonly known as HSV amplicons) are able to transfer genes into both dividing and quiescent cells, and thus have the potential to be widely used as vectors in physiological studies and gene therapy. Historically, these vectors were produced by superinfection with a helper virus that furnished all the trans-acting functions required for amplification and packaging of vector genomes into HSV-1 particles. In these systems, however, large amounts of potentially harmful helper virus are present in the vector stocks, thus restricting the use of these vectors. New helper virus-free packaging systems have been developed that utilize transfection of helper functions rather than infection and thus produce safer vector stocks. The vector titers as well as the amounts of particles obtained with these systems are, however, limited by the impossibility to reamplify the vector stocks. In this article, we present a novel system for producing large amounts of high-titer amplicon vector with low contamination by helper viruses. This system is based on the use of the Cre-loxP recombination system, which allows efficient deletion of the packaging signal of an HSV-1 recombinant helper virus (HSV-1-LaL) on Cre-expressing cells (TE-CRE30).
辅助依赖型单纯疱疹病毒载体（通常称为单纯疱疹病毒扩增子）能够将基因导入分裂细胞和静止细胞，因此有潜力在生理学研究和基因治疗中广泛用作载体。从历史上看，这些载体是通过辅助病毒的超感染产生的，该辅助病毒提供了将载体基因组扩增和包装到单纯疱疹病毒1型颗粒中所需的所有反式作用功能。然而，在这些系统中，载体储备中存在大量潜在有害的辅助病毒，从而限制了这些载体的使用。已经开发出新型无辅助病毒包装系统，该系统利用辅助功能的转染而非感染，从而产生更安全的载体储备。然而，这些系统获得的载体滴度以及颗粒数量受到无法重新扩增载体储备的限制。在本文中，我们提出了一种新型系统，用于生产大量高滴度扩增子载体，且辅助病毒污染低。该系统基于Cre-loxP重组系统的使用，该系统允许在表达Cre的细胞（TE-CRE30）上有效删除单纯疱疹病毒1型重组辅助病毒（HSV-1-LaL）的包装信号。
