通过体外Cre-lox重组高效生成重组腺病毒载体。
Efficient generation of recombinant adenoviral vectors by Cre-lox recombination in vitro.
作者信息
Aoki K, Barker C, Danthinne X, Imperiale M J, Nabel G J
机构信息
Howard Hughes Medical Institute, University of Michigan Medical Center, Department of Internal Medicine and Biological Chemistry, Ann Arbor 48109-0650, USA.
出版信息
Mol Med. 1999 Apr;5(4):224-31.
BACKGROUND
Although recombinant adenovirus vectors are attractive for use in gene expression studies and therapeutic applications, the construction of these vectors remains relatively time-consuming. We report here a strategy that simplifies the production of adenoviruses using the Cre-loxP system.
MATERIALS AND METHODS
Full-length recombinant adenovirus DNA was generated in vitro by Cre-mediated recombination between loxP sites in a linearized shuttle plasmid containing a transgene and adenovirus genomic DNA.
RESULTS
After transfection of Cre-treated DNA into 293 cells, replication-defective viral vectors were rapidly obtained without detectable wild-type virus.
CONCLUSION
This system facilitates the development of recombinant adenoviral vectors for basic and clinical research.
背景
尽管重组腺病毒载体在基因表达研究和治疗应用中具有吸引力，但这些载体的构建仍然相对耗时。我们在此报告一种使用Cre-loxP系统简化腺病毒生产的策略。
材料和方法
通过Cre介导的含有转基因的线性穿梭质粒中的loxP位点与腺病毒基因组DNA之间的重组，在体外产生全长重组腺病毒DNA。
结果
将经Cre处理的DNA转染到293细胞中后，迅速获得了复制缺陷型病毒载体，未检测到野生型病毒。
结论
该系统有助于开发用于基础和临床研究的重组腺病毒载体。
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