Recombinant adenoviruses are attracting a great deal of attention as a highly efficient gene delivery technology and used for in vitro and in vivo gene expression in science. However, among traditional methods, there have been many difficulties and no simple procedure to generate recombinant adenoviruses. Since almost of all these methods involve the process of homologous recombination in a mammalian packaging cell line, the problems are low efficiency of homologous recombination, the need for complicated techniques and the demand for a long time to generate recombinant viruses. These problems have prevented widespread use of adenovirus technology as effective gene transfer tools. In the last few years, there have been several significant and innovative advances in adenoviral technologies, which include a new generation of vectors, the ease of vector manipulation and improvement of the viral production system with homologous recombination in E. coli. Here, we describe the easier and more efficient viral production systems and provide a practical manual for generating recombinant adenoviruses based on one of such systems.
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