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Journal of Virology, April 2002, p. 3774-3783, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.3774-3783.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Improved Primate Foamy Virus Vectors and Packaging Constructs

Martin Heinkelein,1,2 Marco Dressler,1 Gergely Jármy,1 Matthias Rammling,1 Horst Imrich,1 Jana Thurow,1 Dirk Lindemann,1 and Axel Rethwilm1,2*

Institut für Virologie und Immunbiologie, Universität Würzburg,1 Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus," Technische Universität Dresden, Dresden, Germany2

Received 31 August 2001/ Accepted 15 January 2002

Foamy virus (FV) vectors that have minimal cis-acting sequences and are devoid of residual viral gene expression were constructed and analyzed by using a packaging system based on transient cotransfection of vector and different packaging plasmids. Previous studies indicated (i) that FV gag gene expression requires the presence of the R region of the long terminal repeat and (ii) that RNA from packaging constructs is efficiently incorporated into vector particles. Mutants with changes in major 5' splice donor (SD) site located in the R region identified this sequence element as responsible for regulating gag gene expression by an unidentified mechanism. Replacement of the FV 5' SD with heterologous splice sites enabled expression of the gag and pol genes. The incorporation of nonvector RNA into vector particles could be reduced to barely detectable levels with constructs in which the human immunodeficiency virus 5' SD or an unrelated intron sequence was substituted for the FV 5' untranslated region and in which gag expression and pol expression were separated on two different plasmids. By this strategy, efficient vector transfer was achieved with constructs that have minimal genetic overlap.

* Corresponding author. Mailing address: Institut für Virologie, Medizinische Fakultät "Carl Gustav Carus," Technische Universität Dresden, Fetscherstr. 74, 01307 Dresden, Germany. Phone: (49)-351-458-6200. Fax: (49)-351-458-6314. E-mail: Axel.Rethwilm{at}mailbox.tu-dresden.de.

Journal of Virology, April 2002, p. 3774-3783, Vol. 76, No. 8
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.8.3774-3783.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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