A novel approach for preparation of minicircle HSV amplicons by adenovirus mediated Cre-loxP recombination in mammalian cells

作者：Zhao Jie; Liu Xinjing; Han Zhiqiang; Wang Qingzhi; Xu Yuming^*

来源：Life Science Journal-Acta Zhengzhou University Overseas Edition, 2012, 9(3): 413-420.

• Cre-loxP recombination
• Amplicon
• Mini circle amplicon
• HSV
• Gene transfer
• Adenovirus

Amplicon is a plasmid backbone based helper-dependent pseudovirion vector of HSV-1 and has been used as a powerful and versatile gene delivery vehicle due to its unique features. However, like plasmid-based vectors, the major drawback of conventional amplicon for gene delivery is its transient transgene expression, which has been demonstrated that the bacteria elements in the vectors are responsible for the transgene loss event, as evidenced, minicircle DNA and minicircle amplicon devoid of bacteria sequences mediated higher and sustained gene expression both in vitro and in vivo. Nevertheless, current techniques for MC DNA preparation by inducing MC producer plasmid intra-molecular recombination in bacteria have critical limitations, including their labor-intensive, time-consuming procedure, and high contamination with input plasmids and mini plasmids. We thus herein described a novel simple approach for MC-amplicon preparation by utilizing adenovirus mediated Cre-loxP site-specific recombination and HSV helper virus supplied replication and package function in mammalian cells. This technique allows for production of MC-amplicons free of bacteria elements, making it feasible to use MC amplicon instead of conventional amplicon in gene delivery studies. [Jie Zhao, Xinjing Liu, Zhiqiang Han, Qingzhi Wang, Yuming Xu. A novel approach for preparation of minicircle HSV amplicons by adenovirus mediated Cre-loxP recombination in mammalian cells. Life Sci J 2012; 9(3): 413-420]. (ISSN: 1097-8135). http://www.lifesciencesite.com.

• 出版日期2012
• 单位郑州大学