Cre levels limit packaging signal excision efficiency in the Cre/loxP helper-dependent adenoviral vector system

Journal of Virology
Philip NgFrank L Graham

Abstract

Helper-dependent (HD) adenovirus vectors devoid of all viral coding sequences have a large cloning capacity and provide long-term transgene expression in vivo with negligible toxicity, making them attractive vectors for gene therapy. Currently, the most efficient means of producing HD vectors involves coinfecting 293 cells expressing Cre with the HD vector and a helper virus bearing a packaging signal flanked by loxP sites. Cre-mediated packaging signal excision renders the helper virus genome unpackageable but still able to replicate and provide helper functions for HD vector propagation. Typically, helper virus contamination is < or =1% pre- and < or =0.1% postpurification by CsCl banding. While these contamination levels are low, further reduction is desirable. However, this objective has not been realized since the Cre/loxP system was first developed. This lack of progress is due, at least in part, to our lack of understanding of the origins of the contaminating helper virus, thus rendering its reduction or elimination difficult to achieve. This study was designed to investigate the possible sources of contaminating helper virus persisting during HD vector amplification. The results revealed that Cre is limiting in helper v...Continue Reading

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Citations

Apr 29, 2011·Gene Therapy·M Gonzalez-AparicioR Hernandez-Alcoceba
Mar 18, 2003·Human Gene Therapy·Kiran SakhujaSheila Connelly
Feb 11, 2005·Human Gene Therapy·Donna J Palmer, Philip Ng
Jan 2, 2003·The Journal of Physiology·Federica del Monte, Roger J Hajjar
Sep 9, 2010·Expert Opinion on Biological Therapy·Sai V Vemula, Suresh K Mittal
Sep 29, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Donna J Palmer, Philip Ng
Nov 6, 2003·Molecular Therapy : the Journal of the American Society of Gene Therapy·Donna Palmer, Philip Ng
Aug 1, 2010·Viruses·Imre Kovesdi, Susan J Hedley
Dec 23, 2003·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Claire SoudaisEric J Kremer
May 28, 2020·International Journal of Molecular Sciences·Ana RicobarazaRuben Hernandez-Alcoceba
Jul 5, 2011·Cold Spring Harbor Protocols·Donna J Palmer, Philip Ng

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