单纯疱疹病毒1型质粒载体无辅助病毒转移至神经细胞
Helper virus-free transfer of herpes simplex virus type 1 plasmid vectors into neural cells.
作者信息
Fraefel C, Song S, Lim F, Lang P, Yu L, Wang Y, Wild P, Geller A I
机构信息
Division of Endocrinology, Childrens's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
出版信息
J Virol. 1996 Oct;70(10):7190-7. doi: 10.1128/JVI.70.10.7190-7197.1996.
Abstract
Herpes simplex virus type 1 (HSV-1) plasmid vectors have promise for genetic intervention in the brain, but several problems caused by the helper virus have compromised their utility. To develop a helper virus-free packaging system for these vectors, the DNA cleavage/packaging signals were deleted from a set of cosmids that represents the HSV-1 genome. Following cotransfection into cells, this modified cosmid set supported replication and packaging of vector DNA. However, in the absence of the DNA cleavage/packaging signals, the HSV-1 genome was not packaged, and consequently vector stocks were free of detectable helper virus. In the absence of helper virus, the vectors efficiently infected rat neural cells in culture or in the brain with minimal cytopathic effects. beta-galactosidase-positive cells were observed for at least 1 month in vivo, and vector DNA persisted for this period. This system may facilitate studies on neuronal physiology and potential therapeutic applications.
摘要
1型单纯疱疹病毒（HSV-1）质粒载体有望用于脑部的基因干预，但辅助病毒引发的几个问题影响了它们的实用性。为开发针对这些载体的无辅助病毒包装系统，从一组代表HSV-1基因组的黏粒中删除了DNA切割/包装信号。将这种修饰后的黏粒组共转染到细胞中后，它支持载体DNA的复制和包装。然而，在没有DNA切割/包装信号的情况下，HSV-1基因组未被包装，因此载体储备中没有可检测到的辅助病毒。在没有辅助病毒的情况下，这些载体能有效感染培养的或脑内的大鼠神经细胞，且细胞病变效应最小。在体内观察到β-半乳糖苷酶阳性细胞至少持续1个月，并且在此期间载体DNA持续存在。该系统可能有助于神经元生理学研究和潜在的治疗应用。
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