CHAPTER 8 Use of Recombinant Adenovirus for Metabolic Engineering of Mammalian Cells Thomas C. Becker, Richard J. Noel, Ward S. Coats, Anna M. Gomez-Foix,* Tausif Alam, Robert D. Gerard, and Christopher B. Newgard Departments of Biochemistry and Internal Medicine and Gifford Laboratories for Diabetes Research University of Texas Southwestern Medical Center Dallas, Texas 75235 1. 11. 111. IV. V. VI. VII. Historical Perspective on the Development and Application of Viral Vectors for DNA Transfer Examples of the Utility of Recombinant Adenovirus for Studies on Metabolic Regulation A. Pioneering Studies B. Adenovirus-Mediated Gene Transfer to Mammalian Cells in Culture C. In Vivo Studies Adenovirus Biology Development of Adenovirus Vectors Specific Procedures for Preparation of Recombinant Viruses A. Purification of Plasmid DNA B. Co-transfection of 293 Cells C. Extraction of Viral DNA from 293 Cells D. Isolation and Amplification of Viral Clones E. Infection of Mammalian Cells with Recombinant Virions in Vitro F. Delivery of Recombinant Virions in Vivo Future Directions Conclusions References * Current Address: Departament de Bioquimica i Fisiologia, Facultat de Quimica, Universitat de Barcelona, 08028 Barcelona, Spain. METHODS IN CELL BIOLOGY. VOL. 43 Copyright 0 1994 by Academic Press. Inc.. All rights of reproduction in any form reserved 161