利用昆虫病毒系统生产表达胰岛素的腺相关病毒

腺相关病毒(Adeno-associated virus-2,AAV-2)用于构建基因治疗病毒载体,在基因治疗领域受到了普遍重视和关注,而rAAV在应用过程中一个很重要的限制因素就是缺乏简单有效的大规模制备方法.运用昆虫细胞表达系统来提高AAV-2的产量,分别采用绿色荧光蛋白(GFP)和胰岛素(Insulin)基因替代AAV-2的结构基因后,与其他重组病毒共转染293t细胞,3 d后即可检测到高效表达的GFP和Insulin.结果表明,该方法能够比较理想地提高AAV的产量,为临床使用奠定了基础.

Production of Insulin-expressing Adeno-associated Virus(AAV) Using Baculovirus System

As a viral vector of gene therapy,adeno-associated virus-2(AAV-2) has been widely used.But one of the most limitations for the application of AAV-2 is the absence of a simple large-scale production method.We utilized the insect baculovirus system to produce AAV-2 carrying GFP and Insulin express cassette.Expressing of GFP and insulin were detected after 3 days post infection to HEK 293t cells.This method could increase the production of AAV,which could be further used for clinical application.