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

Jinyong Luo, Zhong Liang Deng, Xiaoji Luo, Ni Tang, Wen Xin Song, Jin Chen, Katie A. Sharff, Hue H. Luu, Rex C. Haydon, Kenneth W Kinzler, Bert Vogelstein, Tong Chuan He

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Pages (from-to)1236-1247
Number of pages12
JournalNature Protocols
Volume2
Issue number5
DOIs
StatePublished - May 2007

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Adenoviridae
Plasmids
Viruses
Gene expression
Bacteria
Packaging
Genes
Homologous Recombination
Cells
Eukaryotic Cells
Product Packaging
Transfection
Testing
Gene Expression
Cell Line
Therapeutics

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Luo, J., Deng, Z. L., Luo, X., Tang, N., Song, W. X., Chen, J., ... He, T. C. (2007). A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. Nature Protocols, 2(5), 1236-1247. https://doi.org/10.1038/nprot.2007.135

A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. / Luo, Jinyong; Deng, Zhong Liang; Luo, Xiaoji; Tang, Ni; Song, Wen Xin; Chen, Jin; Sharff, Katie A.; Luu, Hue H.; Haydon, Rex C.; Kinzler, Kenneth W; Vogelstein, Bert; He, Tong Chuan.

In: Nature Protocols, Vol. 2, No. 5, 05.2007, p. 1236-1247.

Research output: Contribution to journalArticle

Luo, J, Deng, ZL, Luo, X, Tang, N, Song, WX, Chen, J, Sharff, KA, Luu, HH, Haydon, RC, Kinzler, KW, Vogelstein, B & He, TC 2007, 'A protocol for rapid generation of recombinant adenoviruses using the AdEasy system', Nature Protocols, vol. 2, no. 5, pp. 1236-1247. https://doi.org/10.1038/nprot.2007.135
Luo, Jinyong ; Deng, Zhong Liang ; Luo, Xiaoji ; Tang, Ni ; Song, Wen Xin ; Chen, Jin ; Sharff, Katie A. ; Luu, Hue H. ; Haydon, Rex C. ; Kinzler, Kenneth W ; Vogelstein, Bert ; He, Tong Chuan. / A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. In: Nature Protocols. 2007 ; Vol. 2, No. 5. pp. 1236-1247.
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