A protocol for rapid generation of recombinant adenoviruses using the AdEasy system

J Luo, ZL Deng, X Luo, N Tang, WX Song, J Chen… - Nature protocols, 2007 - nature.com
J Luo, ZL Deng, X Luo, N Tang, WX Song, J Chen, KA Sharff, HH Luu, RC Haydon
Nature protocols, 2007nature.com
Recombinant adenoviruses provide a versatile system for gene expression studies and
therapeutic applications. We have developed an approach that simplifies the generation and
production of such viruses called the AdEasy system. A recombinant adenoviral plasmid is
generated with a minimum of enzymatic manipulations, employing homologous
recombination in bacteria rather than in eukaryotic cells. After transfection of such plasmids
into a mammalian packaging cell line, viral production is conveniently followed with the aid …
Abstract
Recombinant adenoviruses provide a versatile system for gene expression studies and therapeutic applications. We have developed an approach that simplifies the generation and production of such viruses called the AdEasy system. A recombinant adenoviral plasmid is generated with a minimum of enzymatic manipulations, employing homologous recombination in bacteria rather than in eukaryotic cells. After transfection of such plasmids into a mammalian packaging cell line, viral production is conveniently followed with the aid of GFP encoded by a gene incorporated into the viral backbone. This system has expedited the process of generating and testing recombinant adenoviruses for a variety of purposes. In this protocol, we describe the practical aspects of using the AdEasy system for generating recombinant adenoviruses. The full protocol usually takes 4–5 weeks to complete.
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