An adenovirus type 5 mutant with the preterminal protein gene deleted efficiently provides helper functions for the production of recombinant adeno-associated virus.
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, Ad5dl308DeltapTP was as efficient as the phenotypically wild-type Ad5dl309 in permitting growth of rAAV. Use of Ad5dl308DeltapTP, 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 Ad5dl308DeltapTP or Ad5dl309 as a helper demonstrated that the heat inactivation protocol generally used does not remove all of the helper Ad5dl309 function.
Maxwell
I H
IH
Department of Dermatology, University of Colorado Health Sciences Center, Denver, Colorado 80262, USA.
Maxwell
F
F
Schaack
J
J
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D002460:Cell Line; D003673:Defective Viruses / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D017353:Gene Deletion; D006801:Humans; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics*; D011498:Protein Precursors / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics*
D010750:Phosphoproteins; D011498:Protein Precursors; D014764:Viral Proteins; C085476:pTP protein, adenovirus