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JoVE Journal Bioengineering
Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5
 

Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5

Article DOI: 10.3791/52894-v • 13:17 min • January 28th, 2016
January 28th, 2016 •
1, 1, 1, 1, 1, 1, 1, 1, 1
1Institute of Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, University Witten/Herdecke

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Summary
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A protocol for generation of high-capacity adenoviral vectors lacking all viral coding sequences is presented. Cloning of transgenes contained in the vector genome is based on homing endonucleases. Virus amplification in producer cells grown as adherent cells and in suspension relies on a helper virus providing viral genes in trans.

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Keywords: Adenoviral Vector High-capacity Gene Therapy Transgene Delivery Gene Correction Gene Addition Vector Amplification Vector Purification Helper Virus Cell Culture Freeze/thaw
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