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Journal of Virology, September 1998, p. 7653-7658, Vol. 72, No. 9
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Lipofection of Purified Adeno-Associated Virus Rep68 Protein: toward a Chromosome-Targeting Nonviral Particle

Stefania Lamartina, Giuseppe Roscilli, Daniela Rinaudo, Paola Delmastro, and Carlo Toniatti*

Department of Genetics, Istituto di Ricerche di Biologia Molecolare, I.R.B.M.---Piero Angeletti, 00040 Pomezia, Rome, Italy

Received 5 February 1998/Accepted 3 June 1998

Adeno-associated virus (AAV) integrates very efficiently into a specific site (AAVS1) of human chromosome 19. Two elements of the AAV genome are sufficient: the inverted terminal repeats (ITRs) and the Rep78 or Rep68 protein. The incorporation of the AAV integration machinery in nonviral delivery systems is of great interest for gene therapy. We demonstrate that purified recombinant Rep68 protein is functionally active when directly delivered into human cells by using the polycationic liposome Lipofectamine, promoting the rescue-replication of a codelivered ITR-flanked cassette in adenovirus-infected cells and its site-specific integration in noninfected cells. The sequencing of cloned virus-host DNA junctions confirmed that lipofected Rep68 protein triggers site-specific integration at the same sites in chromosome 19 already characterized in cells latently infected with AAV.

* Corresponding author. Mailing address: Istituto di Ricerche di Biologia Molecolare, I.R.B.M.---P.Angeletti, Via Pontina Km 30,600, 00040 Pomezia, Rome, Italy. Phone: 39-6-91093668. Fax: 39-6-91093654. E-mail: toniatti{at}irbm.it.

Journal of Virology, September 1998, p. 7653-7658, Vol. 72, No. 9
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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