Better Gene Expression by (-)Gene than by (+)Gene in Phage Gene Delivery Systems

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• 作者：Yun Liang ; Bizhi Shi ; Jie Zhang ; Hua Jiang ; Yuhong Xu ; Zonghai Li ; and Jianren Gu
• 刊名：Biotechnology Progress
• 出版年：2006
• 出版时间：June 2006
• 年：2006
• 卷：22
• 期：3
• 页码：626 - 630
• 全文大小：195K
• 年卷期：v.22,no.3(June 2006)
• ISSN：1520-6033

文摘

In recent years, capsid-modified filamentous bacteriophage has become a potential vector forgene delivery into mammalian cells. However, little was known about how the carried gene inthe single-stranded genome expressed in mammalian cells. To explore whether the orientationof the carried gene affects its expression in the cells, we prepared EGF-displayed phagemidparticles whose genome carried the GFP gene or luciferase gene. The phagemid carried reportergenes either in the same orientation (called (+)gene) or in the contrary orientation (called (-)gene)to filamentous origin. Using these phagemid particles to infect H1299 cells, we found that thephages with (-) reporter genes had about 2-fold transduction efficiency as those with (+) reportergenes. These results indicated that phagemid carrying (-)gene of interest presented a betterprocedure in phage-mediated gene therapy. Furthermore, camptothecin (CPT) treatment wasalso applied and found to enhance both kinds of phagemid particles, and (-)gene still producedabout 1.5- to 2-fold transduction efficiency compared to those with (+)gene. Thus, it is imperativethat we clone the genes of interest in the reverse orientation to filamentous origin to enhancetheir expressions when preparing phagemid gene delivery vectors. Also, the results suggestedthat CPT could enhance both the replication of single-stranded DNA and its transcription.