Gene expression studies and gene therapy require efficient gene delivery into cells. Different technologies by viral and non-viral mechanisms have been used for gene delivery into cells. Small gene vectors transfer across the cell membrane with a relatively high efficiency, but not large genes or entire loci spanning several kilobases, which do not remain intact following introduction. Previously, we developed an efficient delivery system based on herpes virus simplex type 1 (HSV-1) amplicons to transfer large fragments of DNA incorporated in human artificial chromosome (HAC) vectors into the nucleus of human cells. The HSV-1 amplicon lacks the signals for cleavage and replication of its own genome, yet each amplicon has the capacity to incorporate up to 150 kb of exogenous DNA. In this study, we investigated whether the capacity of gene delivery could be increased by simultaneously introducing multiple HSV-1 modified amplicons carrying a gene expressing HAC vector into cells with the aim of generating a single artificial chromosome containing the desired genes. Following co-transduction of two HSV-1 HAC amplicons, artificial chromosomes were successfully generated containing the introduced genes, which were appropriately expressed in different human cell types.

中文翻译：

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单纯疱疹病毒1扩增子的多基因人类人工染色体载体传递。

基因表达研究和基因治疗需要有效的基因传递到细胞中。通过病毒和非病毒机制的不同技术已用于将基因传递到细胞中。小型基因载体以相对较高的效率跨细胞膜转移，但跨过几千个碱基的大型基因或整个基因座则没有，导入后不能保持完整。以前，我们开发了一种基于单纯疱疹病毒1型（HSV-1）扩增子的高效递送系统，可将人类人工染色体（HAC）载体中掺入的DNA大片段转移至人类细胞核中。HSV-1扩增子缺乏用于其自身基因组的切割和复制的信号，但每个扩增子均具有整合多达150 kb外源DNA的能力。在这个研究中，我们研究了通过同时携带多个携带表达HAC载体的基因的HSV-1修饰的扩增子进入细胞，以产生包含所需基因的单个人工染色体的方法，是否可以提高基因传递的能力。共转导两个HSV-1 HAC扩增子后，成功地产生了人工染色体，其中包含引入的基因，这些基因可以在不同的人类细胞类型中适当表达。