Russell D W, Miller A D, Alexander I E
Fred Hutchinson Cancer Research Center, Seattle, WA 98104.
Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8915-9. doi: 10.1073/pnas.91.19.8915.
Vectors based on adeno-associated virus can stably transfer genes by chromosomal integration in recipient cells. In this study we have infected stationary and dividing primary human fibroblast cultures with adeno-associated virus vectors encoding alkaline phosphatase and neomycin phosphotransferase. We find that the transduction frequency of S phase cells is about 200 times that of non-S phase cells. However, neither S phase nor mitosis is essential for transduction. Single-stranded vector genomes survive in stationary cultures and can be recruited for transduction by stimulating these cultures to divide. Stable transductants contain randomly integrated vector sequences. These findings have important implications for the use of adeno-associated virus vectors in gene therapy.
基于腺相关病毒的载体可通过在受体细胞中进行染色体整合来稳定转移基因。在本研究中,我们用编码碱性磷酸酶和新霉素磷酸转移酶的腺相关病毒载体感染了静止和正在分裂的原代人成纤维细胞培养物。我们发现S期细胞的转导频率约是非S期细胞的200倍。然而,S期和有丝分裂对于转导都不是必需的。单链载体基因组在静止培养物中存活,并且可以通过刺激这些培养物分裂而被募集用于转导。稳定的转导子含有随机整合的载体序列。这些发现对于腺相关病毒载体在基因治疗中的应用具有重要意义。