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Journal of Virology, October 1998, p. 8371-8373, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

An Adenovirus Type 5 Mutant with the Preterminal Protein Gene Deleted Efficiently Provides Helper Functions for the Production of Recombinant Adeno-Associated Virus

Ian H. Maxwell,1,2 Francoise Maxwell,1 and Jerome Schaack2,3,4,5,*

Department of Dermatology,1 University of Colorado Cancer Center,2 Molecular Biology Program,3 Biomedical Sciences Program,4 and Department of Microbiology,5 University of Colorado Health Sciences Center, Denver, Colorado 80262

Received 10 April 1998/Accepted 10 June 1998

Production of recombinant adeno-associated virus (rAAV) requires helper functions that have routinely been provided by infection of the producer cells with adenovirus. Complete removal and/or inactivation of progeny adenovirus, present in such rAAV preparations, presents significant difficulty. Here, we report that an adenovirus type 5 (Ad5) mutant with the preterminal protein (pTP) gene deleted can provide helper function for the growth of rAAV. At high multiplicity, Ad5dl308Delta pTP was as efficient as the phenotypically wild-type Ad5dl309 in permitting growth of rAAV. Use of Ad5dl308Delta pTP, which is incapable of replication in the absence of complementation for pTP, as a helper avoids the need to remove contaminating adenovirus infectious activity by heat inactivation or by purification. Comparison of the transducing ability of rAAV generated with either Ad5dl308Delta pTP or Ad5dl309 as a helper demonstrated that the heat inactivation protocol generally used does not remove all of the helper Ad5dl309 function.

* Corresponding author. Mailing address: Department of Microbiology, University of Colorado Health Sciences Center, 4200 E. Ninth Ave., Denver, CO 80262. Phone: (303) 315-6883. Fax: (303) 315-6785. E-mail: jerry.schaack{at}uchsc.edu.

Journal of Virology, October 1998, p. 8371-8373, Vol. 72, No. 10
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.




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