Sun M, Zhang G R, Yang T, Yu L, Geller A I
Division of Endocrinology, Children's Hospital, Boston, MA 02115, USA.
Hum Gene Ther. 1999 Aug 10;10(12):2005-11. doi: 10.1089/10430349950017365.
A helper virus-free herpes simplex virus type 1 (HSV-1) plasmid vector system has potential for both gene therapy and physiological studies, but relatively low titers have complicated use of this system. In this article, the packaging efficiency was improved by optimizing the packaging protocol and by adding noninfectious HSV-1-related particles, i.e., previral DNA replication enveloped particles (PREPs), during the packaging procedure. PREPs contain many of the tegument proteins that are thought to enhance an HSV-1 infection. Use of both the optimized packaging protocol and the PREPs resulted in an approximately 50-fold increase in the titer, and five different HSV-1 vectors were packaged using this procedure. A purified vector stock (7.8x10(8) infectious vector particles/ml) was microinjected into the striatum, the rats were sacrificed 4 days after gene transfer, and the brains were found to contain an average of approximately 6740 X-Gal-positive striatal cells. This improved packaging procedure may augment use of this vector system.
一种无辅助病毒的单纯疱疹病毒1型（HSV-1）质粒载体系统在基因治疗和生理学研究方面都具有潜力，但相对较低的滴度使该系统的应用变得复杂。在本文中，通过优化包装方案以及在包装过程中添加非感染性HSV-1相关颗粒，即前病毒DNA复制包膜颗粒（PREPs），提高了包装效率。PREPs包含许多被认为能增强HSV-1感染的被膜蛋白。优化后的包装方案和PREPs的使用使滴度提高了约50倍，并且使用该程序包装了五种不同的HSV-1载体。将纯化的载体原液（7.8x10(8)个感染性载体颗粒/毫升）显微注射到纹状体中，基因转移4天后处死大鼠，发现大脑中平均约有6740个X-Gal阳性纹状体细胞。这种改进的包装程序可能会增加该载体系统的应用。
